Supplemental Table I. The gene expression of 7 acyltransferases in MOVAS-1 cells. Gene Name (pg)/18s(ng) Gene Name (pg)/18s(ng) Acat1 19.53 Gpat 9.75 Acat. Lclat1 1.5 Agpat1 3.31 Lpcat1 8.15 Agpat.16 Lpcat. Agpat3 71.11 Lpcat3 3.71 Agpat 9.9 Lpcat 9.9 Agpat5 3.3 Lpgat1 9.5 Awat1 N.D. Mboat1 35.55 Awat N.D. Mboat N.D. Dgat1. Mboat7 N.D. Dgat.1 Mogat1 N.D. Gpat1 13.1 Mogat 1.9 Gpat N.D. Sptlc1.1 Gpat3.16 The quantitative expression values of 7 acyltransferase were calculated using the absolute standard curve method of real-time qpcr with plasmid templates containing each full length target gene.
Supplemental Table II. shrna Clone IDs used for generating stable knockdown cells Gene Name Clone ID Acat1 Acat Agpat1 Agpat Agpat3 Agpat Agpat5 Atf6 Dgat1 Dgat Gpat1 Gpat3 Gpat Ire1 Lipin Lpcat1 Lpcat Lpcat3 Lpcat Lpgat1 Mboat1 Mogat Scd Sptlc1 VLMM_3791 V3LMM_3666 TRCN17386 VLMM_696 V3LMM_683 V3LMM_6166 V3LMM_1599 V3LMM_5916 TRCN1789 TRCN1576 TRCN1156 TRCN9985 V3LMM_195 V3LMM_599 TRCN988 TRCN183 V3LMM_5893 V3LMM_51111 V3LMM_519 TRCN1569 V3LMM_5116 TRCN18 V3LMM_51655 TRCN13 MOVAS-1 cells were infected with lentiviruses expressing Acat (1 or ), Agpat (1,, 3,, or 5), Atf6, Dgat(1 or ), Gpat (1, 3 or ), Ire1, Lipin, Lpcat (1,, 3, or ), Lpgat1, Mboat1, Mogat, Scd or Sptlc1 shrna and selected with puromycin for 7 days. Each mrna level was measured by real-time qpcr. Changes in gene expression are reported as ratios relative to the control empty shrna.
Supplemental Table III. Fold changes of lipid contents in VSMCs treated with SCD inhibitor. Rank Fold change (vs. ) p value 1 PA-18:/18: 56.1. PA-16:/16: 3.6. 3 PA-16:/18: 3.. PA-1:/18: 5.5. 5 LPA-18: 9.13. 6 LPA-16: 7.81. 7 DAG-16:/18: 7.1. 8 DAG-16:/16: 6.9.1 9 PE-16:1/: 5.11. 1 PE-16:/:5.68.3 11 LPS-18:.67.3 1 DAG-16:1/18:.18. 13 DAG-18:/18:.7. 1 DAG-1:/18: 3.85. 15 LPS-16: 3.78.8 16 PE-16:/16:1 3.77. 17 PE-16:/18:3 3.7.9 18 PS-17:/17:6 3.68.1 19 PS-18:1/: 3.63.15 PS-16:1/18: 3.1.1 1 LPA-16:1 3.3.7 PE-16:/:6 3.6.18 3 TAG-18:/18:/18: 3.1.1 PE-18:3/: 3.1.1 5 LPA-:5.93.3 6 PE-:/:.86. 7 PE-16:/18:.69. 8 PG-16:/:3.69. 9 DAG-18:/18:1.6.1 3 PG-16:/16:.8.8 31 PG-16:/:1.8.13 3 PE-16:1/18:.6. 33 PS-16:1/18:6.37. 3 PE-18:/:5.37.1 35 TAG-16:/16:/18:.3. 36 PG-16:/16:.9.1 37 PE-16:/:.6.6 38 LPC-16:e..1 39 PE-16:/18:.18.3 PE-16:1/16:1.17.5 1 PG-16:/:1..19
DAG-1:/18:1 1.98.3 3 LPA-18:1 1.93.1 PG-16:1/: 1.93. 5 PS-18:1/:6 1.9.7 6 LPS-:6 1.88. 7 PE-18:/:5 1.86. 8 PS-18:1/:3 1.83.17 9 LPS-16:1 1.79.7 5 PS-16:/18:6 1.76.6 51 LPC-18: 1.76.7 5 PI-18:/18: 1.75.19 53 DAG-16:/18:1 1.73.7 5 PA-18:/: 1.7.18 55 PG-16:16:1 1.71.8 56 CER-18: 1.7. 57 DAG-16:/:3 1.66.3 58 PE-16:/18:1 1.63.15 59 PE-18:/18: 1.6.15 6 LPA-:6 1.59.37 61 LPS-18:1 1.58.7 6 PE-16:1/: 1.58.1 63 PS-18:/:5 1.55. 6 PE-18:/:3 1.5.19 65 SM-: 1.51.9 66 TAG-16:/16:/16: 1.51.9 67 PA-16:/18:1 1.5.1 68 PC-36: 1.9.3 69 PC-36:7 1.5.3 7 CE-:6 1..8 71 PE-16:1/:5 1..7 7 PE-18:1/18: 1.3.1 73 LPA-18: 1.3.33 7 PE-18:/:6 1..15 75 PE-18:/18:3 1.39.1 76 PE-16:1/18:1 1.37.5 77 PI-18:/: 1.36.9 78 PS-:1/: 1.35.35 79 PS-16:/18: 1.3.3 8 PA-16:/16:1 1.3.7 81 CER-: 1.3.1 8 PI-16:/16: 1.33.35 83 PA-16:/:1 1.3.3 8 PS-18:1/:5 1.3.18 85 PE-16:1/18: 1.6.33
86 CE-: 1.6.8 87 CER-: 1.5.16 88 LPS-18: 1..37 89 PE-16:/:3 1..17 9 PE-18:/:5 1.1.1 91 PE-18:1/: 1.1.5 9 PE-18:/: 1.1.1 93 DAG-16:1/18:1 1..3 9 DAG-18:/: 1.18.5 95 PS-17:/17: 1.18.5 96 PS-16:1/18:1 1.18.1 97 PE-16:/: 1.16.3 98 PE-18:1/:5 1.15.8 99 PE-18:/:1 1.15.36 1 PE-18:/: 1.1.8 11 PE-16:1/: 1.13. 1 PE-16:1/:6 1.1.33 13 PS-:/: 1.1.3 1 LPI-18:1 1.11.39 15 PE-18:1/: 1.1.36 16 PS-18:1/: 1.1.1 17 PE-18:/18: 1.1. 18 PE-18:1/:3 1.9.39 19 PE-1:/18:1 1.9. 11 TAG-16:/17:/18: 1.9.5 111 LPI-18: 1.9. 11 SM-: 1.8.1 113 PE-16:/16: 1.8.39 11 PA-16:1/18: 1.8.3 115 PC-36:1 1.7.35 116 PE-16:1/: 1.6. 117 PA-18:/;3 1.6.6 118 PE-18:3/:1 1.6.6 119 TAG-16:1/16:1/16:1 1.5.7 1 LPA-:3 1.5.8 11 PI-16:/: 1.5.5 1 PE-16:/: 1..6 13 PS-16:/16:1 1.3.8 1 PC-8: 1.3.1 15 PE-18:1/18:1 1.3.6 16 SM-: 1.3.6 17 DAG-18:1/: 1..9 18 PE-18:/:3 1..8 19 DAG-18:1/18:1 1.1.9
13 CER-16: 1..5 131 DAG-16:/18: 1..5 13 LPC-:.99.9 133 PG-16:/18:.98.7 13 LPC-18:1.97.6 135 SM-:1.96.5 136 PE-16:1/:1.96.5 137 PA-18:/18:1.96. 138 SM-:1.95. 139 PS-18:/:3.9.8 1 SM-:1.89.37 11 PA-16:1/18:1.89.5 1 PS-18:/:.88.3 13 PE-18:1/:6.88.11 1 PG-16:/18:1.88. 15 PE-18:1/:5.87.38 16 PE-18:/:.87.9 17 PE-16:/:1.86. 18 PA-18:/18:.85. 19 TAG-16:/16:1/17:.8.36 15 CER-:1.8.9 151 PG-16:/18:.8.31 15 PC-36:8.8.6 153 DAG-18:/18:.83.3 15 PC-3:1.8.16 155 TAG-1:/17:/18:1.8.3 156 PE-:/:6.81.3 157 PG-18:/18:.81.36 158 PG-16:/18:1.81.3 159 PE-18:1/18:3.8.7 16 PG-16:1/18:.8.39 161 PC-3:1.79.8 16 PE-16:1/:.79.31 163 PE-18:1/:.78.1 16 PS-16:/18:.78.31 165 SM-18:.77.7 166 TAG-18:/18:/18:1.76.3 167 DAG-18:1/18:.76.5 168 TAG-16:/18:/18:1.76.7 169 PE-16:/:5.7.3 17 PC-3:3.7.3 171 PC-3:1.73.7 17 PG-16:1/18:1.73.35 173 PG-16:/18:.73.5
17 PS-16:/18:1.7.31 175 PA-18:1/18:.7.31 176 LPC-:.71.6 177 TAG-16:1/16:1/18:.71.7 178 PC-3:.7. 179 LPC-18:e.7.7 18 PE-18:/18:1.69.15 181 LPC-16:1.69. 18 CE-:5.69.37 183 TAG-1:/18:/18:1.68.1 18 PC-3:.68. 185 CE-18:.68.3 186 PC-3:.66. 187 CE-16:.66.7 188 TAG-18:1/18:1/:.66.33 189 TAG-1:/16:/18:.65.1 19 PC-3:.65.1 191 PE-16:1/:3.65.1 19 PG-16:/18:.6.6 193 PC-3:.6. 19 PC-8:1.63. 195 PE-:/:5.63.18 196 PC-3:5.63.3 197 PC-38:.63. 198 TAG-1:/16:/18:1.63.11 199 PE-:1/:.6.16 TAG-16:/17:/18:1.6. 1 PS-16:/18:16.61. PA-18:1/18:1.6.11 3 LPC-16:.6.5 PG-18:1/18:3.6.31 5 LPC-18:.59.7 6 CE-18:.59.1 7 PC-36:6.59.1 8 PC-8:.59. 9 PC-:5.58. 1 PC-:6.58.3 11 PC-36:.57.1 1 PC-36:3.56. 13 PC-33:1.5. 1 PC-3:3.5. 15 LPC-18:3.5.18 16 PC-38:3.5. 17 PC-:.5.
18 TAG-18:/18:/18:.51. 19 PC-38:7.5. TAG-15:/18:1/18:1.9.6 1 LPS-:3.9.7 TAG-15:/16:/18:1.9.16 3 LPC-:1.8.15 CE-18:1.8.11 5 PS-16:1/:.8.18 6 LPC-:.7. 7 TAG-1:1/18:/18:.7.1 8 TAG-1:/16:1/18:1.7.11 9 TAG-1:1/16:1/18:.7.11 3 PC-3:.7.1 31 PC-:7.5. 3 DAG-18:1/:3..5 33 PC-38:6.. 3 PC-38:.3. 35 PC-36:.. 36 LPI-:3.1.18 37 PC-36:5.1.1 38 TAG-1:1/16:/18:1.1.13 39 TAG-18:1/18:1/:6.1.1 PC-38:5.1. 1 CE-16:1..5 TAG-18:/18:1/18:1..6 3 PG-16:1/:1..3 LPI-:.39.16 5 TAG-16:1/16:1/18:1.39. 6 TAG-16:/16:/18:.37. 7 TAG-18:1/18:1/18:1.36.1 8 PG-18:1/18:1.35.8 9 PG-18:/18:.3.13 5 LPI-:3.3.17 51 TAG-16:/16:1/18:1.3. 5 TAG-16:/18:1/18:1.33.5 53 PS-18:/:.33.3 5 TAG-18:1/18:/18:.33.7 55 CE-:3.3.5 56 LPI-:.3.15 57 PG-18:1/18:.3.16 58 TAG-16:/18:/18:.9.6 59 PE-18:/:.8.15 6 LPC-:3.7.3 61 LPI-18:.6.19
6 TAG-16:/18:1/18:.6.7 63 TAG-16:1/17:/18:1.6.5 6 TAG-16:1/18:1/18:.6. 65 TAG-16:1/18:1/18:.6. 66 LPC-:.5.9 67 TAG-18:18:/18:..6 68 TAG-18:1/18:1/18:.3.3 69 LPI-18:.1.18 7 PG-1:/18:1..1 71 PG-1:/18:1..16 7 TAG-17:/18:1/18:1.16.3 73 TAG-17:/18:1/18:1.16.3 7 TAG-1:1/18:1/18:1.16.7 75 TAG-16:1/18:1/18:1.15. 76 TAG-16:/17:/18:.15. 77 PG-16:/:..19 78 PG-16:1/:3..3 79 PG-18:/18:3..19 VSMCs were treated with 3nM CAY1566 for hours. Total lipids were extracted using Bligh & Dyer's method and analyzed with LC-MS/MS. CE; choletseryl ester, DAG; CER; ceramide, diacylglycerol, LPA; lysophosphatidic acid; LPC; lysophosphatidylcholine, LPE; lysophosphatidiyl ethanolamine, LPI; lysophosphatidylinsitol, LPS; lysophosphatidylserinepa; phosphatidic acid, PC; phosphatidylcholine, PE; phosphatidylethanolamine, PG; phosphatidylglycerol PI; phosphatidylinositol, PS; phosphatidylserine, SM; sphingomyelin; TAG; triacylglycerol,
Supplemental Table IV. Fold changes of lipid content in the medail layer of aortas from SMC-Scd1/ KO mice Rank Fold change (vs control) P value 1 PA-18:/18: 63.. PA-16:/18: 11.9. 3 PA-1:/18: 7.71. PA-16:/16: 7.. 5 PG-16:/16:.5.9 6 PG-16:/:1.15.1 7 PE-16:1/:.8.1 8 LPS-16: 3.73. 9 LPA-18: 3.69. 1 LPC-: 3.65. 11 DAG-16:/18: 3.55.1 1 PE-16:1/: 3.35.1 13 LPS-16:1 3.19.18 1 LPI-18: 3.1.5 15 PI-16:/16: 3..7 16 TAG-16:/17:/18:.95.9 17 PG-16:1/18:.93. 18 PS-16:1/18:.75.6 19 PE-:/:.5.3 DAG-16:1/18:.. 1 LPI-18:.39.15 CE-:5.3.9 3 PS-16:/18:6.9.6 LPS-:6.8. 5 PS-16:1/:..8 6 PS-18:/:.17.1 7 DAG-18:/18:1.16. 8 CER-18:.15.1 9 PS-18:1/:.13.18 3 PG-16:/18:1.1.3 31 DAG-1:/18:.9.5 3 PS-18:1/:.5.6 33 PS-16:/18:.. 3 PA-16:/:1..13 35 PG-18:/18: 1.97.18 36 DAG-16:/16: 1.9.1 37 DAG-18:/18: 1.9.9 38 PS-16:/18: 1.86.1 39 PE-18:/:6 1.85.1 LPI-18:1 1.8. 1 LPI-:3 1.83.16 PG-16:16:1 1.8.9 3 PE-18:/: 1.8. PS-18:1/:6 1.81.15
5 LPC-16:e 1.79.1 6 SM-: 1.78. 7 PE-16:/:6 1.75.1 8 TAG-16:/16:/18: 1.75. 9 PE-16:/:1 1.7.1 5 LPE-18: 1.7.1 51 LPA-16: 1.7. 5 TAG-18:/18:/18: 1.69.1 53 PA-16:1/18: 1.69.3 5 PG-16:1/:1 1.63.3 55 PS-16:/16:1 1.63. 56 CER-: 1.61.3 57 CE-18: 1.61.7 58 LPC-18: 1.59.3 59 LPC-16:1 1.58.1 6 PS-:/: 1.56.19 61 PI-18:/18: 1.56.19 6 LPI-: 1.55.1 63 PE-16:/18: 1.5.8 6 LPC-: 1.53.3 65 PE-16:/: 1.5.8 66 DAG-18:1/: 1.9. 67 LPE-18:1 1.6.3 68 PS-18:/: 1..7 69 CE-16: 1..19 7 PE-18:/18: 1.1. 71 SM-16: 1.1.7 7 PS-18:1/:5 1.1.6 73 PE-16:1/:3 1.1.19 7 PE-16:1/18: 1..9 75 PG-16:/18: 1.39.36 76 LPA-:3 1.38.39 77 PC-36: 1.36.1 78 PI-16:/: 1.3.36 79 PE-16:/16: 1.3.15 8 PE-18:/:5 1.3.15 81 PE-18:1/:5 1.9. 8 PE-18:1/:6 1.9.6 83 PE-18:/: 1.8.13 8 PE-18:/:3 1.8.3 85 SM-18: 1.8.6 86 TAG-16:/16:/16: 1.7.33 87 PA-18:/18:1 1.7.16 88 SM-: 1.7. 89 PG-16:/18: 1.5.1 9 PI-18:/: 1..5 91 LPC-: 1.1.5
9 PE-18:/18:1 1..1 93 PE-16:1/: 1..33 9 PE-18:/18:3 1..33 95 PS-16:1/18:1 1.19.36 96 PC-36:7 1.19.5 97 DAG-18:/18: 1.19.8 98 PE-16:1/:5 1.18.3 99 PE-16:/18:1 1.17.1 1 CE-:3 1.17.3 11 PE-16:/16:1 1.16.3 1 PE-18:/:5 1.16.1 13 LPC-18:e 1.15.3 1 LPS-18: 1.1.35 15 SM-:1 1.13.9 16 PE-16:/:3 1.1.37 17 PA-18:/18: 1.1.3 18 SM-18:1 1.1. 19 PE-18:/:5 1.1.33 11 CE-: 1.1.35 111 SM-: 1.11.18 11 PE-16:/18: 1.9.33 113 PE-16:1/:1 1.8.9 11 PC-36:1 1.7.3 115 SM-:1 1.7.7 116 LPC-18:1 1.7.3 117 PE-16:/: 1.7.6 118 PE-18:/:3 1.6.1 119 PC-8: 1.6.5 1 PE-18:1/18:1 1.6. 11 PA-16:/18:1 1.6.38 1 LPC-16: 1.6.1 13 PE-18:1/:5 1.6.6 1 DAG-1:/18:1 1.5.38 15 TAG-1:/18:/18:1 1..6 16 LPC-18:3 1..7 17 PE-:/:6 1.3.8 18 LPA-18: 1..9 19 PE-18:/: 1..9 13 PI-18:/;3 1..5 131 PA-18:/:3 1..5 13 DAG-18:/: 1..9 133 PS-16:/18:1 1..5 13 TAG-16:/18:/18:1.99.5 135 PE-18:/:.98.9 136 PG-16:1/18:1.98.9 137 PS-16:/18:1.98.9 138 SM-:1.97.1
139 PC-36:8.93.9 1 PE-16:1/:.93. 11 PG-16:/:3.9.7 1 PS-18:1/:3.9.5 13 CE-18:.91. 1 DAG-16:/18:1.89. 15 PS-18:/:3.89.3 16 PE-18:1/:.87.3 17 PE-16:1/18:.86.15 18 CE-:6.86.38 19 CER-:.85.8 15 PE-:/:5.85. 151 PA-18:/:.8.6 15 SM-18:.83.17 153 PE-18:3/:.83.38 15 PE-16:/:.8.9 155 TAG-16:/16:1/17:.81.3 156 LPS-18:1.8.6 157 PS-:1/:.8.39 158 PE-16:/18:3.79.6 159 PE-18:1/:.79.6 16 PE-1:/18:1.78.3 161 PG-18:1/18:1.78.37 16 PS-16:1/18:.78. 163 PE-18:1/:3.77.1 16 LPC-:3.77.16 165 PG-18:1/18:.76.37 166 CE-18:1.76.18 167 PE-18:/18:.76.37 168 LPE-16:.7. 169 PE-16:1/:6.7.3 17 PS-16:/16:.73.37 171 PS-18:/:5.73.3 17 PE-16:1/:.7.3 173 LPC-:1.71.16 17 PC-3:1.71.1 175 PC-3:1.7. 176 PE-18:3/:1.7.1 177 TAG-18:/18:/18:1.69.17 178 PE-18:1/18:3.69. 179 PA-18:1/18:.69.1 18 PE-16:1/18:1.69.3 181 PE-18:1/:.68.9 18 CER-:1.68.3 183 DAG-16:1/18:1.68. 18 PC-3:1.67.3 185 PC-3:.67.5
186 PE-18:1/18:.66.5 187 PE-:1/:.65.9 188 PC-3:3.65.3 189 TAG-18:/18:1/18:1.65.16 19 PC-8:.63.1 191 PC-:6.6. 19 DAG-18:1/18:.6.1 193 CER-16:.6.15 19 PC-3:.61. 195 PC-36:.6. 196 PC-3:.59. 197 DAG-16:/18:.59.7 198 PC-3:.59. 199 PE-18:/:1.59.1 PC-38:.59. 1 TAG-16:1/16:1/18:.58.1 PC-3:.58. 3 DAG-18:1/18:1.57. PC-3:5.57. 5 TAG-16:/17:/18:1.56.15 6 PC-33:1.56. 7 PC-:5.56. 8 TAG-1:/16:/18:1.56.8 9 LPA-18:1.56.3 1 PC-36:3.55. 11 PC-8:1.55. 1 PA-18:1/18:1.55. 13 DAG-16:/:3.5.8 1 LPS-18:.51.18 15 PC-3:3.5. 16 PC-38:6.9. 17 TAG-17:/18:1/18:1.9. 18 TAG-17:/18:1/18:1.9. 19 PC-36:6.8. LPS-:3.7. 1 PE-16:/:5.6.13 PC-38:3.6. 3 PC-36:5.6. TAG-1:1/18:/18:.5.1 5 LPA-:5.5.8 6 LPC-18:..6 7 TAG-15:/16:/18:1..1 8 TAG-1:1/16:1/18:..1 9 LPA-16:1.3.3 3 TAG-1:/16:/18:.3.1 31 PC-38:7.3. 3 PC-38:5..
33 PG-16:1/:3..1 3 DAG-18:1/:3..1 35 PC-:7.. 36 TAG-16:1/17:/18:1..6 37 PG-18:/18:..1 38 PE-16:1/16:1..1 39 TAG-16:/18:/18:..1 TAG-16:/16:/18:.1.1 1 PC-38:.. PC-:.39. 3 CE-16:1.39. TAG-1:1/16:/18:1.37.11 5 PC-36:.36. 6 TAG-1:/17:/18:1.36.9 7 TAG-16:1/16:1/18:1.3.1 8 TAG-16:/17:/18:.31. 9 PA-16:1/18:1.31.1 5 PC-3:.31. 51 PG-1:/18:1.9.11 5 PG-18:1/18:3.8.1 53 TAG-16:1/18:1/18:.7.7 5 TAG-16:1/18:1/18:.7.7 55 LPC-:.7.1 56 TAG-16:/18:1/18:1.7. 57 TAG-16:/18:1/18:.6. 58 TAG-18:1/18:1/:6..1 59 TAG-16:/16:1/18:1..1 6 TAG-18:18:/18:..1 61 TAG-18:1/18:1/18:.3.3 6 TAG-1:/16:1/18:1.3. 63 TAG-18:1/18:/18:.19.5 6 TAG-16:1/18:1/18:1.19.1 65 TAG-18:1/18:1/:.18.7 66 TAG-18:1/18:1/18:1.18.1 67 TAG-15:/18:1/18:1.16. 68 TAG-16:1/16:1/16:1.13.5 69 TAG-1:1/18:1/18:1.13. 7 TAG-18:/18:/18:.11. 71 PE-16:/:5.1.5 7 LPA-:6..19 73 CE-:1..7 Mice were sacrificed at 18-week-old. Total lipids in the aortic media were extracted by Bligh & Dyer's Method and were analyzed with LC-MS/MS. CE; choletseryl ester, DAG; CER; ceramide, diacylglycerol, LPA; lysophosphatidic acid; LPC; lysophosphatidylcholine, LPE; lysophosphatidiyl ethanolamine, LPI; lysophosphatidylinsitol, LPS; lysophosphatidylserinepa; phosphatidic acid, PC; phosphatidylcholine, PE; phosphatidylethanolamine, PG; phosphatidylglycerol
Supplemental Table V. qpcr primer list Gene Primer Bank ID Forward Reverse Acat1 639a1 GAAACCGGCTGTCAAAATCTGG TGTGACCATTTCTGTATGTGTCC Acat 157a1 ACAAGACAGACCTCTTCCCTC ATGGTTCGGAAATGTTGCACC Agpat1 6359a1 TAAGATGGCCTTCTACAACGGC CCATACAGGTATTTGACGTGGAG Agpat 395616a1 CAGCCAGGTTCTACGCCAAG TGATGCTCATGTTATCCACGGT Agpat3 7978a1 CTGCTTGCCTACCTGAAGACC GATACGGCGGTATAGGTGCTT Agpat 796a1 CCAGTTTCTATGTCACCTGGTC GCAGAGTCTGGCATTGATCTTG Agpat5 7977a1 CACACGTACTCTATGCGCTAC AAGAAGAGCACCATGTTCTGG Alp 6671533a1 CCAACTCTTTTGTGCCAGAGA GGCTACATTGGTGTTGAGCTTTT Atf3 31515a1 GAGGATTTTGCTAACCTGACACC TTGACGGTAACTGACTCCAGC Atf 675318a1 ATGGCGCTCTTCACGAAATC ACTGGTCGAAGGGGTCATCAA Awat1 1877c3 AGAGTGTGCCTAACACCACC CCGAACGTGAAGGTAGGGAC Awat 93a1 TTGACTGGAAGACCCCTGAG TCACCATCTTGAGTGGGAAGTA Chop 319815a1 CTGGAAGCCTGGTATGAGGAT CAGGGTCAAGAGTAGTGAAGGT Dgat1 675363a1 TCCGTCCAGGGTGGTAGTG TGAACAAAGAATCTTGCAGACGA Dgat 169759a1 GCGCTACTTCCGAGACTACTT GGGCCTTATGCCAGGAAACT Gadd3 6678978a1 GAGGGACGCCCACAACTTC TTACCAGAGACAGGGGTAGGT Gpat1 66857a1 ACAGTTGGCACAATAGACGTTT CCTTCCATTTCAGTGTTGCAGA Gpat 63319a1 CACTGCTCCGAGGTTTTGATG AGGTTGGCAGCAATTCCATAC Gpat3 7376a1 GGCCTTCGGATTATCCCTGG CTTGGGGGCTCCTTTCTGAA Gpat 3531a1 AGCTTGATTGTCAACCTCCTG CCGTTGGTGTAGGGCTTGT GPAT 5151c GGTTAGGAGTGCTGATTCGGT CCAGAAGGCTAATCCCTGTGAA Lclat1 9578999c1 TTTTATGCTCGGCCCCATTTT CACAAGACGGCTGCTAATCCA Lipin1 793911a1 CTCCGCTCCCGAGAGAAAG TCATGTGCAAATCCACGGACT Lipin 315319a1 GAAGTGGCGGCTCTCTATTTC AGAGGGTTACATCAGGCAAGT LIPIN 769c1 TCTACAAGGGCATTAACCAGGC AACGTGAAAAGGTGAACACTGA Lipin3 15897a1 CAAACCTCGTGGTGAAAATCAAC CCACAGTGCTCTCAGGTAAGT Lpcat1 315186a1 GGCTCCTGTTCGCTGCTTT TTCACAGCTACACGGTGGAAG Lpcat 7375a1 CCCTTCGTCCAGCAGACTAC GCAGCAAAATTATTCCAACCAGT Lpcat3 169958a1 GACGGGGACATGGGAGAGA GTAAAACAGAGCCAACGGGTAG Lpcat 796911c3 CTCATCCGATACCCCAACAGT GGGAGGAACTCTACATCCACG Mboat1 395631a1 AGCCTCTCTTACCGTACCACC GGCTGGCTTTACCAGGATGTA Mboat 189991a1 TCAGACACGTAGTTGCTACCC TGCAGTAGGAAATCCCACTTTG Mboat7 3151a1 TACCGCACCTACCTGGATTG AGAAGACCGGGATCATGTAGAA Mogat1 18685a1 TGGTGCCAGTTTGGTTCCAG TGCTCTGAGGTCGGGTTCA Mogat 89798a1 TGGGAGCGCAGGTTACAGA CAGGTGGCATACAGGACAGA Ocn N/A GCAATAAGGTAGTGAACAGACTCC GTTTGTAGGCGGTCTTCAAGC Opn 6678113a1 AGCAAGAAACTCTTCCAAGCAA GTGAGATTCGTCAGATTCATCCG PiT1 7657579a1 TTTGACAAACTTCCTCTGTGGG GGACTTTCAGACGGACTAGACTT SCD 5375915c1 TCTAGCTCCTATACCACCACCA TCGTCTCCAACTTATCTCCTCC Scd1 3153675a1 TTCTTGCGATACACTCTGGTGC CGGGATTGAATGTTCTTGTCGT Scd 6677863a1 GCATTTGGGAGCCTTGTACG AGCCGTGCCTTGTATGTTCTG Scd3 1377368a1 GTTGCCACTTTACTGAGATACGC GAAGCCCTCGCCCATACTT Scd 3179a1 GCCCACTTGCCACAAGAGAT GTAGCTGGGGTCATACAGATCA SCD5 1859696c1 TGCGACGCCAAGGAAGAAAT CCTCCAGACGATGTTCTGCC Sm 675571a1 CAACAAGGGTCCATCCTACGG ATCTGGGCGGCCTACATCA Smmhc 19578a1 GAGCAAACTCAGGAGAGGAAAC GTCCCGAGCGTCCATTTCTTC
Sptlc1 9577a1 ACGAGGCTCCAGCATACCAT TCAGAACGCTCCTGCAACTTG Sptlc 6755656a1 AACGGGGAAGTGAGGAACG CAGCATGGGTGTTTCTTCAAAAG sxbp-1 N/A GAGTCCGCAGCAGGTG GTGTCAGAGTCCATGGGA TetR N/A TGCTTAATGAGGTCGGAATCG ATCTCAATGGCTAAGGCGTC The primers for Ocn, sxbp-1 and TetR were designed in the laboratory.
Supplemental Table VI. 3-Qtrpa spectrometer settings for lipidomic analysis Lipid Name Q1 m/z Q3 m/z DP CE EP CXP CEP 1:.3895 369.3 3 5 7 CE 1:1.37335 369.3 3 5 7 CE 15: 68.63 369.3 3 5 7 CE 15:1 66.5873 369.3 3 5 7 CE 16: 6.6189 369.3 3 5 7 CE 16:1 6.63 369.3 3 5 7 CE 17: 656.635 369.3 3 5 7 CE 17:1 65.6186 369.3 3 5 7 CE 18: 67.65 369.3 3 5 7 CE 18:1 668.635 369.3 3 5 7 CE 18:1 669.635 37.3 3 5 7 CE 18: 666.6189 369.3 3 5 7 CE 18: 668.6 371.3 3 5 7 CE 18:3 66.63 369.3 3 5 7 CE 18:3 665.63 37.3 3 5 7 CE : 698.6815 369.3 3 5 7 CE :1 696.6658 369.3 3 5 7 CE : 69.65 369.3 3 5 7 CE :3 69.635 369.3 3 5 7 CE : 69.6189 369.3 3 5 7 CE : 69.6189 369.3 3 5 7 CE : 69.6189 371.3 3 5 7 CE :5 688.63 369.3 3 5 7 CE :5 67.63 371.3 3 5 7 CE : 76.718 369.3 3 5 7 CE :1 7.6969 369.3 3 5 7 CE : 7.6815 369.3 3 5 7 CE : 718.65 369.3 3 5 7 CE :5 716.65 369.3 3 5 7 CE :6 71.6183 369.3 3 5 7 LPC 13: 5.3 18.1 5 35 5 3 LPC 13: 55.3 185.1 5 35 5 3 LPC 16:e 8.3 18.1 5 35 5 3 LPC 16:1 9.3 18.1 5 35 5 3 LPC 16: 96.3 18.1 5 35 5 3 LPC 16: 97.3 185.1 5 35 5 3 LPC 18: 5.3 18.1 5 35 5 3 LPC 18:3 518.3 18.1 5 35 5 3 LPC 18:1 5. 18.1 5 35 5 3 LPC 18: 5. 18.1 5 35 5 3 LPC 18:e 51.3 18.1 5 35 5 3 LPC : 55. 18.1 5 35 5 3 LPC o-: 538. 1.1 5 35 5 3 LPC : 58. 18.1 5 35 5 3 LPC o-: 566.5 1.1 5 35 5 3
LPC :1 578. 18.1 5 35 5 3 LPC :3 56. 18.1 5 35 5 3 LPC : 5.3 18.1 5 35 5 3 LPC :5 5.3 18.1 5 35 5 3 LPC :6 568.3 18.1 5 35 5 3 LPC : 68.5 18.1 5 35 5 3 LPC o-: 59.5 1.1 5 35 5 3 LPC o-:1 59.5 1.1 5 35 5 3 LPC o-: 59.5 1.1 5 35 5 3 LPS 13: 56. 71. 5 5 5 16 LPS 16:1 96.3 311.3 5 5 5 16 LPS 16: 98.3 313.3 5 5 5 16 LPS 18: 5.3 337.3 5 5 5 16 LPS 18:1 5.3 339.3 5 5 5 16 LPS 18: 56.3 31.3 5 5 5 16 LPS :3 58.3 33.3 5 5 5 16 LPS : 56.3 31.3 5 5 5 16 LPS :5 5.3 339.3 5 5 5 16 LPS :6 57.3 31.3 5 5 5 16 LPG 13: 3.3 5.3 8 15 LPG 16:1 83.3 9.3 8 15 LPG 16: 85.3 96.3 8 15 LPG 18: 59.3 3.3 8 15 LPG 18:1 511.3 3.3 8 15 LPG 18: 513.3 3.3 8 15 LPG :3 535.3 36.3 8 15 LPG : 533.3 3.3 8 15 LPG :5 531.3 3.3 8 15 LPG :6 557.3 368.3 8 15 LPI 13: 58 1 18 3 3 3 LPI 16:1 586 1 18 3 3 3 LPI 16: 588 1 18 3 3 3 LPI 18: 61 1 18 3 3 3 LPI 18:1 616 1 18 3 3 3 LPI 18: 618 1 18 3 3 3 LPI :3 6 1 18 3 3 3 LPI : 638 1 18 3 3 3 LPI :5 636 1 18 3 3 3 LPI :6 66 1 18 3 3 3 LPI 13: 58 71 1 3 3 3 LPI 16:1 586 39 1 3 3 3 LPI 16: 588 311 1 3 3 3 LPI 18: 61 337 1 3 3 3 LPI 18:1 616 339 1 3 3 3 LPI 18: 618 31 1 3 3 3 LPI :3 6 363 1 3 3 3 LPI : 638 361 1 3 3 3
LPI :5 636 359 1 3 3 3 LPI :6 66 385 1 3 3 3 LPA 16:1 6. 155 5 3 1 LPA 16: 8. 155 5 3 1 LPA 17:.3 155 5 3 1 LPA 18: 5.3 155 5 3 1 LPA 18:1 5.3 155 5 3 1 LPA 18: 56.3 155 5 3 1 LPA :3 68.3 155 5 3 1 LPA : 76.3 155 5 3 1 LPA :5 5.3 155 5 3 1 LPA :6 83.3 155 5 3 1 LPE 13: 1. 71. 5 3 15 LPE 16:e.3 99.3 5 3 15 LPE 16:1 5.3 311.3 5 3 15 LPE 16: 5.3 313.3 5 3 15 LPE 18: 78.3 337.3 5 3 15 LPE 18:3 76.3 335.3 5 3 15 LPE 18:1 8.3 339.3 5 3 15 LPE 18: 8.3 31.3 5 3 15 LPE 18:e 68.3 37.3 5 3 15 LPE : 51.3 369.3 5 3 15 LPE :3 5.3 363.3 5 3 15 LPE : 5.3 361.3 5 3 15 LPE :5 5.3 359.3 5 3 15 LPE :6 53.3 389.3 5 3 15 CER 1: 8.6 6. 35 5 3 5 CER 16: 538.7 6. 35 5 3 5 CER 18: 566.7 6. 35 5 3 5 CER : 59.7 6. 35 5 3 5 CER :1 68.9 6. 35 5 3 5 CER : 65.9 6. 35 5 3 5 DHCer 16: 5.7 66. 35 5 3 5 DHCer 18: 568.7 66. 35 5 3 5 DHCer :1 65.9 66. 35 5 3 5 DHCer : 65.9 66. 35 5 3 5 GlcCre 1: 6.6 6. 35 5 5 3 5 GlcCre 16: 7.7 6. 35 5 5 3 5 GlcCre 18: 78.7 6. 35 5 5 3 5 GlcCre : 756 6. 35 5 5 3 5 GlcCre : 78 6. 35 5 5 3 5 GlcCre :1 81.9 6. 35 5 5 3 5 GlcCre : 81 6. 35 5 5 3 5 Cer1P 1: 56.6 6. 5 5 3 5 Cer1P 16: 618.7 6. 5 5 3 5 Cer1P : 73.9 6. 5 5 3 5 DHCer1P 1: 56.6 66. 5 5 3 5
DHCer1P : 73.9 66. 5 5 3 5 SM 1: 67.7 18.1 55 5 3 3 SM 16: 73. 18.1 55 5 3 3 SM 18: 731. 18.1 55 5 3 3 SM 18:1 79. 18.1 55 5 3 3 SM 18: 77. 18.1 55 5 3 3 SM :1 757. 18.1 55 5 3 3 SM : 759. 18.1 55 5 3 3 SM :1 785. 18.1 55 5 3 3 SM : 788 18.1 55 5 3 3 SM :1 813. 18.1 55 5 3 3 SM : 815 18.1 55 5 3 3 DHSM 18: 733.8 18.1 55 5 3 3 DHSM : 817.9 18.1 55 5 3 3 PC : 6.5 18.1 6 7 3 3 PC 6: 65.5 18.1 6 7 3 3 PC 8: 678.5 18.1 6 7 3 3 PC 8:1 675.5 18.1 6 7 3 3 PC 8: 67.5 18.1 6 7 3 3 PC 3: 76.5 18.1 6 7 3 3 PC 3:1 7.5 18.1 6 7 3 3 PC 3: 7.5 18.1 6 7 3 3 PC 3: 73.5 18.1 6 7 3 3 PC 3:1 73.5 18.1 6 7 3 3 PC 3: 73.5 18.1 6 7 3 3 PC 3:3 78.5 18.1 6 7 3 3 PC 33:1 76.5 18.1 6 7 3 3 PC 3: 76.5 18.1 6 7 3 3 PC 3:1 76.5 18.1 6 7 3 3 PC 3: 758.5 18.1 6 7 3 3 PC 3:3 756.5 18.1 6 7 3 3 PC 3: 75.5 18.1 6 7 3 3 PC 3:5 75.5 18.1 6 7 3 3 PC 36: 79.5 18.1 6 7 3 3 PC 36:1 788.5 18.1 6 7 3 3 PC 36: 786.5 18.1 6 7 3 3 PC 36:3 78.5 18.1 6 7 3 3 PC 36: 78.5 18.1 6 7 3 3 PC 36:5 78.5 18.1 6 7 3 3 PC 36:6 778.5 18.1 6 7 3 3 PC 36:7 776.5 18.1 6 7 3 3 PC 36:8 77.5 18.1 6 7 3 3 PC 38: 81.5 18.1 6 7 3 3 PC 38:3 81.5 18.1 6 7 3 3 PC 38: 81.5 18.1 6 7 3 3 PC 38:5 88.5 18.1 6 7 3 3 PC 38:6 86.5 18.1 6 7 3 3
PC 38:7 8.5 18.1 6 7 3 3 PC : 838.5 18.1 6 7 3 3 PC :5 836.5 18.1 6 7 3 3 PC :6 838.5 18.1 6 7 3 3 PC :7 836.5 18.1 6 7 3 3 PC o-3: 69.5 18.1 6 7 3 3 PC o-3: 7.5 18.1 6 7 3 3 PC o-3:1 718.5 18.1 6 7 3 3 PC o-3: 716.5 18.1 6 7 3 3 PC o-3: 78.5 18.1 6 7 3 3 PC o-3:1 76.5 18.1 6 7 3 3 PC o-3: 7.5 18.1 6 7 3 3 PC o-3:3 7.5 18.1 6 7 3 3 PC o-3: 7.5 18.1 6 7 3 3 PC o-36: 776.5 18.1 6 7 3 3 PC o-36:1 77.5 18.1 6 7 3 3 PC o-36: 77.5 18.1 6 7 3 3 PC o-36:3 77.5 18.1 6 7 3 3 PC o-36: 768.5 18.1 6 7 3 3 PC o-36:5 766.5 18.1 6 7 3 3 PC o-38: 795.3 18.1 6 7 3 3 PC o-38:5 793.5 18.1 6 7 3 3 PC o-38:6 791.5 18.1 6 7 3 3 PC o-:6 8.5 18.1 6 7 3 3 PC o-:7 818.5 18.1 6 7 3 3 PC 3p: 69.5 18.1 6 7 3 3 PC 3p: 718.5 18.1 6 7 3 3 PC 3p:1 716.5 18.1 6 7 3 3 PC 3p: 76.5 18.1 6 7 3 3 PC 3p:1 7.5 18.1 6 7 3 3 PC 3p: 7.5 18.1 6 7 3 3 PC 3p:3 7.5 18.1 6 7 3 3 PC 36p: 77.5 18.1 6 7 3 3 PC 36p:1 77.5 18.1 6 7 3 3 PC 36p: 77.5 18.1 6 7 3 3 PC 36p:3 768.5 18.1 6 7 3 3 PC 36p: 766.5 18.1 6 7 3 3 PC 36p:5 76.5 18.1 6 7 3 3 PC 38p: 79.5 18.1 6 7 3 3 PC 38p:5 79.5 18.1 6 7 3 3 PC 38p:6 79.5 18.1 6 7 3 3 PC p:5 8.5 18.1 6 7 3 3 PC p:6 818.5 18.1 6 7 3 3 PE 17:/17: 7.6 579.6 5 18 5 7 PE 3: 688.5 57.5 5 18 5 7 PE 3:1 69.5 59.5 5 18 5 7 PE 3: 69.5 551.5 5 18 5 7
PE 3: 7.5 579.5 5 18 5 7 PE 3:1 718.5 577.5 5 18 5 7 PE 3: 716.5 575.5 5 18 5 7 PE 3:3 71.5 573.5 5 18 5 7 PE 35:1 73.5 591.5 5 18 5 7 PE 35: 73.5 589.5 5 18 5 7 PE 36:5 738.5 597.5 5 18 5 7 PE 36: 7.5 599.5 5 18 5 7 PE 36:3 7.5 61.5 5 18 5 7 PE 36: 7.5 63.5 5 18 5 7 PE 36:1 76.5 65.5 5 18 5 7 PE 38:7 76.5 61.5 5 18 5 7 PE 38:6 76.5 63.5 5 18 5 7 PE 38:5 766.5 65.5 5 18 5 7 PE 38: 768.5 67.5 5 18 5 7 PE 38.3 77.5 69.5 5 18 5 7 PE :7 79.5 69.5 5 18 5 7 PE :6 79.5 651.5 5 18 5 7 PE :5 79.5 653.5 5 18 5 7 PE : 796.5 655.5 5 18 5 7 PE p:3 81.5 669.5 5 18 5 7 PE p: 81.5 671.5 5 18 5 7 PE :9 81.5 673.5 5 18 5 7 PE :8 816.5 675.5 5 18 5 7 PE :7 818.5 677.5 5 18 5 7 PE :6 8.5 679.5 5 18 5 7 PE 3p: (16:/18:) 7.5 559.5 5 18 5 7 PE 3p:1 (16:/18:1) 7.5 561.5 5 18 5 7 PE 36p: (16:/:) 7.5 583.5 5 18 5 7 PE 36p:3 (16:/:3) 76.5 585.5 5 18 5 7 PE 36p: (18:/18:) 78.5 587.5 5 18 5 7 PE 36p:1 (18:/18:1) 73.5 589.5 5 18 5 7 PE 38p:6 (16:/:6) 78.5 67.5 5 18 5 7 PE 38p:5 (16:/:5) 75.5 69.5 5 18 5 7 PE 38p: (18:/:) 75.5 611.5 5 18 5 7 PE p:6 (18:/:6) 776.5 635.5 5 18 5 7 PE p:5 (18:/:5) 778.5 637.5 5 18 5 7 PE p: (:/:) 78.5 639.5 5 18 5 7 PE 3o:1 7.6 563.6 5 18 5 7 PE 3o: 7.5 561.5 5 18 5 7 PE 36o:1 73.5 591.5 5 18 5 7 PE 36o: 73.5 589.5 5 18 5 7 PE 36o:3 78.6 587.6 5 18 5 7 PE 36o: 76.5 585.5 5 18 5 7 PE 36o:5 7.5 583.5 5 18 5 7 PE 36o:6 7.5 581.5 5 18 5 7 PE 38o: 75.6 613.6 5 18 5 7
PE 38o:5 75.6 611.6 5 18 5 7 PE o:5 78.6 639.6 5 18 5 7 PE o:6 778.5 637.5 5 18 5 7 PE o:7 776.6 635.6 5 18 5 7 PS 3: 736.5 551.5 6 18 7 8 PS 3:1 73.5 59.5 6 18 7 8 PS 3: 76.5 579.5 6 18 7 8 PS 3:1 76.5 577.5 6 18 7 8 PS 36: 788.5 63.5 6 18 7 8 PS 36: 79.5 67.5 6 18 7 8 PS 36:1 79.5 65.5 6 18 7 8 PS 38:5 81.5 65.5 6 18 7 8 PS 38: 81.5 67.5 6 18 7 8 PS 38:3 81.5 69.5 6 18 7 8 PS :7 83.5 69.5 6 18 7 8 PS :6 836.5 651.5 6 18 7 8 PS :5 838.5 653.5 6 18 7 8 PS :8 83.5 67.5 6 18 7 8 PS : 8.5 655.5 6 18 7 8 PI 3: 88.5 18.1 6 7 3 3 PI 3:1 86.5 18.1 6 7 3 3 PI 3: 856.5 18.1 6 7 3 3 PI 3:3 85.5 18.1 6 7 3 3 PI 3: 85.5 18.1 6 7 3 3 PI 3:1 85.5 18.1 6 7 3 3 PI 36: 876.5 18.1 6 7 3 3 PI 36:3 878.5 18.1 6 7 3 3 PI 36: 88.5 18.1 6 7 3 3 PI 36:1 88.5 18.1 6 7 3 3 PI 36: 88.5 18.1 6 7 3 3 PI 38:6 9.5 18.1 6 7 3 3 PI 38:5 9.5 18.1 6 7 3 3 PI 38: 9.5 18.1 6 7 3 3 PI 38:3 96.5 18.1 6 7 3 3 PI 38: 98.5 18.1 6 7 3 3 PI :6 98.5 18.1 6 7 3 3 PI :5 93.5 18.1 6 7 3 3 PI : 93.5 18.1 6 7 3 3 PG 3: 7.5 551.5 5 18 5 8 5 PG 3:1 738.5 59.5 5 18 5 8 5 PG 3: 768.6 579.6 5 18 5 8 5 PG 3: 76.5 575.5 5 18 5 8 5 PG 3:1 766.5 577.5 5 18 5 8 5 PG 36: 788.5 599.5 5 18 5 8 5 PG 36:3 79.5 61.5 5 18 5 8 5 PG 36: 79.5 63.5 5 18 5 8 5 PG 36:1 79.5 65.5 5 18 5 8 5
PA 3:3 66.5 155 35 5 5 3 35 PA 3: 66.5 155 35 5 5 3 35 PA 3:1 66.5 155 35 5 5 3 35 PA 3: 666.5 155 35 5 5 3 35 PA 3: 69.5 155 35 5 5 3 35 PA 3:1 69.5 155 35 5 5 3 35 PA 3: 69.5 155 35 5 5 3 35 PA 36: 718.5 155 35 5 5 3 35 PA 36:1 7.5 155 35 5 5 3 35 PA 36: 7.5 155 35 5 5 3 35 PA 38: 7.5 155 35 5 5 3 35 FFA 1: 18. 18. 5 5 5 7 FFA 13: 3. 3. 5 5 5 7 FFA 1: 6. 6. 5 5 5 7 FFA 15: 6. 6. 5 5 5 7 FFA 16:1 7. 7. 5 5 5 7 FFA 16: 7. 7. 5 5 5 7 FFA 17:1 86.3 86.3 5 5 5 7 FFA 17: 88.3 88.3 5 5 5 7 FFA 18:3 96.3 96.3 5 5 5 7 FFA 18: 98.3 98.3 5 5 5 7 FFA 18:1 3.3 3.3 5 5 5 7 FFA 18: 3.3 3.3 5 5 5 7 FFA 19: 316.3 316.3 5 5 5 7 FFA :5 3.3 3.3 5 5 5 7 FFA : 3.3 3.3 5 5 5 7 FFA :3 3.3 3.3 5 5 5 7 FFA : 36.3 36.3 5 5 5 7 FFA :1 38.3 38.3 5 5 5 7 FFA : 33.3 33.3 5 5 5 7 FFA :6 36.3 36.3 5 5 5 7 FFA :5 38.3 38.3 5 5 5 7 FFA : 35.3 35.3 5 5 5 7 FFA : 35. 35. 5 5 5 7 FFA :1 356. 356. 5 5 5 7 FFA : 358. 358. 5 5 5 7 FFA :1 38. 38. 5 5 5 7 FFA : 386. 386. 5 5 5 7 MAG 1: 33.3 58.1 3 5 8 5 1 MAG 1:1 31.3 75.1 3 5 8 5 1 MAG 15: 317.3 75.1 3 5 8 5 1 MAG 16:1 39.3 75.1 3 5 8 5 1 MAG 16: 331.3 75.1 3 5 8 5 1 MAG 17:1 33.3 75.1 3 5 8 5 1 MAG 17: 35.3 75.1 3 5 8 5 1 MAG 18:3 353.3 75.1 3 5 8 5 1 MAG 18: 355.3 75.1 3 5 8 5 1
MAG 18:1 357.3 75.1 3 5 8 5 1 MAG 18: 359.3 75.1 3 5 8 5 1 MAG 19: 373. 75.1 3 5 8 5 1 MAG :5 377.3 75.1 3 5 8 5 1 MAG : 379.3 75.1 3 5 8 5 1 MAG :3 381.3 75.1 3 5 8 5 1 MAG : 383.3 75.1 3 5 8 5 1 MAG :1 385. 75.1 3 5 8 5 1 MAG : 387. 75.1 3 5 8 5 1 MAG :6 3.3 75.1 3 5 8 5 1 MAG :5 5.3 75.1 3 5 8 5 1 MAG : 3.3 75.1 3 5 8 5 1 MAG : 11. 75.1 3 5 8 5 1 MAG :1 13. 75.1 3 5 8 5 1 MAG : 15. 75.1 3 5 8 5 1 DAG 3: 558.5 313.3 3 3 5 DAG 3:1 58.5 85.3 3 3 5 DAG 3: 58.5 85.3 3 3 5 DAG 3: 558.5 99.3 3 3 5 DAG 3: 586.5 313.3 3 3 5 DAG 3: 61.6 31.3 3 3 5 DAG 3:1 61.6 339.3 3 3 5 DAG 3: 61.6 313.3 3 3 5 DAG 36:3 636.6 313.3 3 3 5 DAG 36: 63.5 313.3 3 3 5 DAG 38:6 66.6 313.3 3 3 5 DAG 38:6 658.5 313.3 3 3 5 DAG 3:1 61.6 311.3 3 3 5 DAG 3: 61.5 339.3 3 3 5 DAG 3: 61.6 37.3 3 3 5 DAG 36: 6.6 31.3 3 3 5 DAG 36:1 6.6 339.3 3 3 5 DAG 36: 638.6 31.3 3 3 5 DAG 38: 66.6 31.3 3 3 5 DAG :6 686.6 31.3 3 3 5 DAG 36: 638.6 339.3 3 3 5 DAG 36:3 636.6 339.3 3 3 5 DAG 36: 63.6 339.3 3 3 5 DAG 38: 66.6 339.3 3 3 5 DAG 38:5 66.6 339.3 3 3 5 DAG :7 68.6 339.3 3 3 5 DAG 36: 63.5 337.3 3 3 5 TAG 8:1 8.8 53.5 6 3 1 6 5 TAG 8: 8.8 57.5 6 3 1 6 5 TAG 8: 8.8 51.5 6 3 1 6 5 TAG 8:3 818.8 51.5 6 3 1 6 5 TAG 9:1 836.8 537.5 6 3 1 6 5
TAG 5:1 85.8 65.6 6 3 1 6 5 TAG 5: 8.8 599.6 6 3 1 6 5 TAG 8: 8.8 577.6 6 3 1 6 5 TAG 8: 8.8 59.5 6 3 1 6 5 TAG 5:3 86.8 63.6 6 3 1 6 5 TAG 5:3 86.8 57.5 6 3 1 6 5 TAG 5: IS 78.7 53. 6 3 1 6 5 TAG 9:1 836.8 577.5 6 3 1 6 5 TAG 51: 86.8 63.5 6 3 1 6 5 TAG 8: 8.8 551.5 6 3 1 6 5 TAG 5: 85.8 551.5 6 3 1 6 5 TAG 5:1 85.8 551.5 6 3 1 6 5 TAG 5: 88.8 551.5 6 3 1 6 5 TAG 9:1 836.8 563.5 6 3 1 6 5 TAG 5: 88.8 59.5 6 3 1 6 5 TAG 51: 866.8 593.6 6 3 1 6 5 TAG 51:1 86.8 565.5 6 3 1 6 5 TAG 51: 86.8 589.6 6 3 1 6 5 TAG 5:1 878.8 577.5 6 3 1 6 5 TAG 5: 876.8 63.6 6 3 1 6 5 TAG 5:3 87.8 577.6 6 3 1 6 5 TAG 5: 87.8 599.6 6 3 1 6 5 TAG 8:3 818.8 57.5 6 3 1 6 5 TAG 5: 88.8 57.5 6 3 1 6 5 TAG 5:3 87.8 576.5 6 3 1 6 5 TAG 51: 86.8 563.5 6 3 1 6 5 TAG 5:3 87.8 63.5 6 3 1 6 5 TAG 5: 87.8 573.6 6 3 1 6 5 TAG 53: 89.8 63.6 6 3 1 6 5 TAG 51: IS 866.8 579.6 6 3 1 6 5 TAG 5: 98.9 67.6 6 3 1 6 5 TAG 5:1 96.9 67.6 6 3 1 6 5 TAG 5: 9.9 63.6 6 3 1 6 5 TAG 5: 9.8 599.5 6 3 1 6 5 TAG 5:3 9.9 63.6 6 3 1 6 5 TAG 5: 9.9 63.7 6 3 1 6 5 TAG 56:6 9.9 63.6 6 3 1 6 5 TAG 58:8 98.9 63.7 6 3 1 6 5 TAG 5:5 898.9 599.6 6 3 1 6 5 TAG 5:6 896.9 599.6 6 3 1 6 5 TAG 56:8 9.9 599.6 6 3 1 6 5 MHC 16: 7.6 6.3 35 5 3 5 MHC 18: 78.6 6.3 35 5 3 5 MHC : 756.6 6.3 35 5 3 5 MHC : 78.7 6.3 35 5 3 5 MHC : 81.7 6.3 35 5 3 5 MHC :1 81.7 6.3 35 5 3 5
DHC 16: 86.6 6.3 35 5 3 5 DHC 18: 89.7 6.3 35 5 3 5 DHC : 918.7 6.3 35 5 3 5 DHC : 96.7 6.3 35 5 3 5 DHC : 97.8 6.3 35 5 3 5 DHC :1 97.7 6.3 35 5 3 5 THC 17: 138.7 6.3 35 5 3 5 THC 16: 1.7 6.3 35 5 3 5 THC 18: 15.7 6.3 35 5 3 5 THC : 18.7 6.3 35 5 3 5 THC : 118.8 6.3 35 5 3 5 THC : 1136.8 6.3 35 5 3 5 THC :1 113.8 6.3 35 5 3 5 GM3 16: 1153.7 6.3 35 5 3 5 GM3 18: 1181.8 6.3 35 5 3 5 GM3 : 19.8 6.3 35 5 3 5 GM3 : 137.8 6.3 35 5 3 5 GM3 : 165.9 6.3 35 5 3 5 GM3 :1 163.8 6.3 35 5 3 5 LPS LPS-13: 5. 153. -3. -35. -6. -5. -18. LPS LPS-16:1 9.3 153. -3. -35. -6. -5. -18. LPS LPS-16: 96.3 153. -3. -35. -6. -5. -18. LPS LPS-18: 5.3 153. -3. -35. -6. -5. -18. LPS LPS-18:1 5.3 153. -3. -35. -6. -5. -18. LPS LPS-18: 5.3 153. -3. -35. -6. -5. -18. LPS LPS-:3 56.3 153. -3. -35. -6. -5. -18. LPS LPS-: 5.3 153. -3. -35. -6. -5. -18. LPS LPS-:5 5.3 153. -3. -35. -6. -5. -18. LPS LPS-:6 568.3 153. -3. -35. -6. -5. -18. LPI LPI-13: 59. 13. -55. -3. -6. -5. -18. LPI LPI-16:1 567. 53. -55. -3. -6. -5. -18. LPI LPI-16: 569. 55. -55. -3. -6. -5. -18. LPI LPI-18: 595. 79. -55. -3. -6. -5. -18. LPI LPI-18:1 597. 81. -55. -3. -6. -5. -18. LPI LPI-18: 599. 83.3-55. -3. -6. -5. -18. LPI LPI-:3 61. 35. -55. -3. -6. -5. -18. LPI LPI-: 619. 33. -55. -3. -6. -5. -18. LPI LPI-:5 617. 31. -55. -3. -6. -5. -18. LPI LPI-:6 63. 37. -55. -3. -6. -5. -18. LPA LPA-16:1 7. 153. -. -3. -6. -5. -18. LPA LPA-16: 9. 153. -. -3. -6. -5. -18. LPA LPA-17: 3.3 153. -. -3. -6. -5. -18. LPA LPA-18: 33.3 153. -. -3. -6. -5. -18. LPA LPA-18:1 35.3 153. -. -3. -6. -5. -18. LPA LPA-18: 37.3 153. -. -3. -6. -5. -18. LPA LPA-:3 59.3 153. -. -3. -6. -5. -18. LPA LPA-: 57.3 153. -. -3. -6. -5. -18.
LPA LPA-:5 55.3 153. -. -3. -6. -5. -18. LPA LPA-:6 6.3 153. -. -3. -6. -5. -18. LPE LPE-13: 1. 13. -35. -3. -6. -5. -18. LPE LPE-16:e.3 99.3-35. -3. -6. -5. -18. LPE LPE-16:1 5.3 53. -35. -3. -6. -5. -18. LPE LPE-16: 5.3 55. -35. -3. -6. -5. -18. LPE LPE-18: 78.3 79. -35. -3. -6. -5. -18. LPE LPE-18:3 76.3 77. -35. -3. -6. -5. -18. LPE LPE-18:1 8.3 81. -35. -3. -6. -5. -18. LPE LPE-18: 8.3 83.3-35. -3. -6. -5. -18. LPE LPE-18:e 68.3 37.3-35. -3. -6. -5. -18. LPE LPE-: 51.3 311.3-35. -3. -6. -5. -18. LPE LPE-:3 5.3 35. -35. -3. -6. -5. -18. LPE LPE-: 5.3 33. -35. -3. -6. -5. -18. LPE LPE-:5 5.3 31. -35. -3. -6. -5. -18. LPE LPE-:6 53.3 37. -35. -3. -6. -5. -18. LPG LPG-13: 1.3 13. -. -35. -6. -5. -18. LPG LPG-16:1 81.3 53. -. -35. -6. -5. -18. LPG LPG-16: 83.3 55. -. -35. -6. -5. -18. LPG LPG-18: 57.3 79. -. -35. -6. -5. -18. LPG LPG-18:1 59.3 81. -. -35. -6. -5. -18. LPG LPG-18: 511.3 83.3 -. -35. -6. -5. -18. LPG LPG-:3 533.3 35. -. -35. -6. -5. -18. LPG LPG-: 531.3 33. -. -35. -6. -5. -18. LPG LPG-:5 59.3 31. -. -35. -6. -5. -18. LPG LPG-:6 555.3 37. -. -35. -6. -5. -18. PE 17:/17: 718.6 69. -6. -5. -6. -5. -18. PE 3: 686.5 53. -6. -5. -6. -5. -18. PE 3: 686.5 79. -6. -5. -6. -5. -18. PE 3:1 688.5 53. -6. -5. -6. -5. -18. PE 3:1 688.5 81. -6. -5. -6. -5. -18. PE 3: 69.5 55. -6. -5. -6. -5. -18. PE 3: 69.5 83.3-6. -5. -6. -5. -18. PE 3: 718.5 83.3-6. -5. -6. -5. -18. PE 3:1 716.5 55. -6. -5. -6. -5. -18. PE 3:1 716.5 53. -6. -5. -6. -5. -18. PE 3: 71.5 79. -6. -5. -6. -5. -18. PE 3: 71.5 81. -6. -5. -6. -5. -18. PE 3:3 71.5 77. -6. -5. -6. -5. -18. PE 3:3 71.5 53. -6. -5. -6. -5. -18. PE 36:5 736.5 31. -6. -5. -6. -5. -18. PE 36:5 736.5 53. -6. -5. -6. -5. -18. PE 36:5 736.5 77. -6. -5. -6. -5. -18. PE 36: 738.5 33. -6. -5. -6. -5. -18. PE 36: 738.5 53. -6. -5. -6. -5. -18. PE 36: 738.5 77. -6. -5. -6. -5. -18. PE 36: 738.5 79. -6. -5. -6. -5. -18.
PE 36:3 7.5 35. -6. -5. -6. -5. -18. PE 36:3 7.5 37.3-6. -5. -6. -5. -18. PE 36:3 7.5 77. -6. -5. -6. -5. -18. PE 36:3 7.5 81. -6. -5. -6. -5. -18. PE 36: 7.5 37.3-6. -5. -6. -5. -18. PE 36: 7.5 53. -6. -5. -6. -5. -18. PE 36: 7.5 79. -6. -5. -6. -5. -18. PE 36: 7.5 81. -6. -5. -6. -5. -18. PE 36:1 7.5 39.3-6. -5. -6. -5. -18. PE 36:1 7.5 53. -6. -5. -6. -5. -18. PE 36:1 7.5 81. -6. -5. -6. -5. -18. PE 38:7 76.5 37. -6. -5. -6. -5. -18. PE 38:7 76.5 31. -6. -5. -6. -5. -18. PE 38:7 76.5 77. -6. -5. -6. -5. -18. PE 38:6 76.5 37. -6. -5. -6. -5. -18. PE 38:6 76.5 81. -6. -5. -6. -5. -18. PE 38:6 76.5 33. -6. -5. -6. -5. -18. PE 38:6 76.5 77. -6. -5. -6. -5. -18. PE 38:5 76.5 39. -6. -5. -6. -5. -18. PE 38:5 76.5 331.3-6. -5. -6. -5. -18. PE 38:5 76.5 31. -6. -5. -6. -5. -18. PE 38:5 76.5 81. -6. -5. -6. -5. -18. PE 38:5 76.5 35. -6. -5. -6. -5. -18. PE 38:5 76.5 77. -6. -5. -6. -5. -18. PE 38: 766.5 331.3-6. -5. -6. -5. -18. PE 38: 766.5 33. -6. -5. -6. -5. -18. PE 38: 766.5 81. -6. -5. -6. -5. -18. PE 38: 766.5 37.3-6. -5. -6. -5. -18. PE 38: 766.5 77. -6. -5. -6. -5. -18. PE 38.3 768.5 335.3-6. -5. -6. -5. -18. PE 38.3 768.5 35. -6. -5. -6. -5. -18. PE 38.3 768.5 81. -6. -5. -6. -5. -18. PE 38.3 768.5 39.3-6. -5. -6. -5. -18. PE 38.3 768.5 77. -6. -5. -6. -5. -18. PE :7 788.5 37. -6. -5. -6. -5. -18. PE :7 788.5 331.3-6. -5. -6. -5. -18. PE :7 788.5 35. -6. -5. -6. -5. -18. PE :6 79.5 37. -6. -5. -6. -5. -18. PE :6 79.5 81. -6. -5. -6. -5. -18. PE :6 79.5 331.3-6. -5. -6. -5. -18. PE :5 79.5 39. -6. -5. -6. -5. -18. PE :5 79.5 81. -6. -5. -6. -5. -18. PE :5 79.5 31. -6. -5. -6. -5. -18. PE :5 79.5 33. -6. -5. -6. -5. -18. PE : 79.5 331.3-6. -5. -6. -5. -18. PE : 79.5 33. -6. -5. -6. -5. -18. PE :9 81.5 37. -6. -5. -6. -5. -18.
PE :8 81.5 331.3-6. -5. -6. -5. -18. PE :8 81.5 37. -6. -5. -6. -5. -18. PE :6 818.5 37. -6. -5. -6. -5. -18. PE PE-3p: (16:/18:) 698.5 79. -6. -5. -6. -5. -18. PE PE-3p:1 (16:/18:1) 7.5 81. -6. -5. -6. -5. -18. PE PE-3p:1 (16:/18:) 7.5 83.3-6. -5. -6. -5. -18. PE PE-36p: (16:/:) 7.5 33. -6. -5. -6. -5. -18. PE PE-36p:3 (16:/:3) 7.5 35. -6. -5. -6. -5. -18. PE PE-36p: (18:/18:) 76.5 79. -6. -5. -6. -5. -18. PE PE-36p:1 (18:/18:1) 78.5 81. -6. -5. -6. -5. -18. PE PE-36p:1 (18:/18:) 73.5 83.3-6. -5. -6. -5. -18. PE PE-38p:6 (16:/:6) 76.5 37. -6. -5. -6. -5. -18. PE PE-38p:5 (16:/:5) 78.5 39. -6. -5. -6. -5. -18. PE PE-38p: (18:/:) 75.5 331.3-6. -5. -6. -5. -18. PE PE-p:6 (18:/:6) 77.5 37. -6. -5. -6. -5. -18. PE PE-p:5 (18:/:5) 776.5 39. -6. -5. -6. -5. -18. PE PE-p: (:/:) 778.5 33. -6. -5. -6. -5. -18. PS PS-16:/16: 736.5 55. -6. -58. -6. -5. -18. PS PS-1:/18: 736.5 83.3-6. -58. -6. -5. -18. PS PS-16:/16:1 73.5 53. -6. -58. -6. -5. -18. PS PS-1:/18:1 73.5 81. -6. -58. -6. -5. -18. PS PS-16:/18: 76.5 83.3-6. -58. -6. -5. -18. PS PS-17:/17: 76.5 69. -6. -58. -6. -5. -18. PS PS-16:/18:1 76.5 81. -6. -58. -6. -5. -18. PS PS-16:/18:1 76.5 55. -6. -58. -6. -5. -18. PS PS-16:1/18: 76.5 53. -6. -58. -6. -5. -18. PS PS-16:1/18: 76.5 83. -6. -58. -6. -5. -18. PS PS-16:/18: 788.5 79. -6. -58. -6. -5. -18. PS PS-16:/18: 788.5 55. -6. -58. -6. -5. -18. PS PS-16:1/18:1 788.5 53. -6. -58. -6. -5. -18. PS PS-16:/18: 79.5 83.3-6. -58. -6. -5. -18. PS PS-17:/17: 79.5 69. -6. -58. -6. -5. -18. PS PS-16:/18:1 79.5 81. -6. -58. -6. -5. -18. PS PS-16:/18:1 79.5 55. -6. -58. -6. -5. -18. PS PS-16:1/18: 79.5 53. -6. -58. -6. -5. -18. PS PS-16:/:6 88.5 37. -6. -58. -6. -5. -18. PS PS-16:/:5 81.5 39. -6. -58. -6. -5. -18. PS PS-16:1/: 81.5 331.3-6. -58. -6. -5. -18. PS PS-18:1/: 81.5 33. -6. -58. -6. -5. -18. PS PS-18:/:3 81.5 79. -6. -58. -6. -5. -18. PS PS-16:/: 81.5 55. -6. -58. -6. -5. -18. PS PS-18:/: 81.5 33. -6. -58. -6. -5. -18. PS PS-18:/: 81.5 83. -6. -58. -6. -5. -18. PS PS-18:1/:3 81.5 81. -6. -58. -6. -5. -18. PS PS-18:/:3 81.5 35. -6. -58. -6. -5. -18. PS PS-18:1/:6 83.5 81. -6. -58. -6. -5. -18. PS PS-18:/:6 836.5 37. -6. -58. -6. -5. -18.
PS PS-18:1/:5 836.5 39. -6. -58. -6. -5. -18. PS PS-18:/: 836.5 331.3-6. -58. -6. -5. -18. PS PS-18:/:5 838.5 39. -6. -58. -6. -5. -18. PS PS-18:1/; 838.5 81. -6. -58. -6. -5. -18. PS PS-:/:5 838.5 31. -6. -58. -6. -5. -18. PS PS-;1/: 838.5 39.3-6. -58. -6. -5. -18. PS PS-18:/:6 83.5 37. -6. -58. -6. -5. -18. PS PS-18:3/:5 83.5 77. -6. -58. -6. -5. -18. PS PS-:/: 83.5 33. -6. -58. -6. -5. -18. PS PS-18:/: 8.5 331.3-6. -58. -6. -5. -18. PS PS-:/: 8.5 33. -6. -58. -6. -5. -18. PS PS-:1/:3 8.5 39.3-6. -58. -6. -5. -18. PI PI-16:/16: 89.5 55. -55. -3. -6. -5. -18. PI PI-18:/18: 865.6 83.3-55. -3. -6. -5. -18. PI PI-16:/: 857.5 33. -55. -3. -6. -5. -18. PI PI-:/18: 98.5 311.3-55. -3. -6. -5. -18. PI PI-18:/:3 887.6 35. -55. -3. -6. -5. -18. PI PI-18:/: 885.5 33. -55. -3. -6. -5. -18. PI PI-16:/:6 9.5 37. -55. -3. -6. -5. -18. PI PI-:/: 95.5 331.3-55. -3. -6. -5. -18. PI PI-18:/:6 99.5 33. -55. -3. -6. -5. -18. PI PI-18:/:5 911.6 39. -55. -3. -6. -5. -18. PI PI-18:/: 913.6 83.3-55. -3. -6. -5. -18. PG PG-1:/18: 71.5 83.3-7. -5. -6. -5. -18. PG PG-16:/16: 71.5 55. -7. -5. -6. -5. -18. PG PG-1:/18:1 719.5 81. -7. -5. -6. -5. -18. PG PG-16:/16:1 719.5 53. -7. -5. -6. -5. -18. PG PG-16:/18: 79.5 55. -7. -5. -6. -5. -18. PG PG-17:/17: 79.5 69. -7. -5. -6. -5. -18. PG PG-16:/18: 75.5 79. -7. -5. -6. -5. -18. PG PG-16:1/18:1 75.5 81. -7. -5. -6. -5. -18. PG PG-16:/18:1 77.5 55. -7. -5. -6. -5. -18. PG PG-16:1/18: 77.5 83.3-7. -5. -6. -5. -18. PG PG-16:/: 769.5 33. -7. -5. -6. -5. -18. PG PG-16:1/:3 769.5 53. -7. -5. -6. -5. -18. PG PG-18:1/18:3 769.5 77. -7. -5. -6. -5. -18. PG PG-18:/18: 769.5 79. -7. -5. -6. -5. -18. PG PG-16:/:3 771.5 35. -7. -5. -6. -5. -18. PG PG-18:/18:3 771.5 83.3-7. -5. -6. -5. -18. PG PG-18:1/18: 771.5 79. -7. -5. -6. -5. -18. PG PG-16:1/:1 773.5 39. -7. -5. -6. -5. -18. PG PG-18:/18: 773.5 79. -7. -5. -6. -5. -18. PG PG-18:1/18:1 773.5 81. -7. -5. -6. -5. -18. PG PG-16:/:1 775.5 39. -7. -5. -6. -5. -18. PG PG-16:1/: 775.5 53. -7. -5. -6. -5. -18. PA PA-1:/18:3 61.1 77. -6. -8. -6. -5. -18. PA PA-1:/18: 63.1 79. -6. -8. -6. -5. -18.
PA PA-1:/18:1 65.1 81. -6. -8. -6. -5. -18. PA PA-16:/16:1 65.1 53. -6. -8. -6. -5. -18. PA PA-1:/18: 67.1 7. -6. -8. -6. -5. -18. PA PA-16:/16: 67.1 55. -6. -8. -6. -5. -18. PA PA-16:/18: 671.1 79. -6. -8. -6. -5. -18. PA PA-16:1/18:1 671.1 53. -6. -8. -6. -5. -18. PA PA-16:1/18: 673.1 53. -6. -8. -6. -5. -18. PA PA-16:/18:1 673.1 81. -6. -8. -6. -5. -18. PA PA-1:/: 675.1 311.3-6. -8. -6. -5. -18. PA PA-16:/18: 675.1 55. -6. -8. -6. -5. -18. PA PA-17:/17: 675.1 69. -6. -8. -6. -5. -18. PA PA-16:/: 699.1 37.3-6. -8. -6. -5. -18. PA PA-16:1/:1 699.1 53. -6. -8. -6. -5. -18. PA PA-18:/18: 699.1 79. -6. -8. -6. -5. -18. PA PA-18:1/18:1 699.1 81. -6. -8. -6. -5. -18. PA PA-16:/:1 71.1 39.3-6. -8. -6. -5. -18. PA PA-16:1/: 71.1 53. -6. -8. -6. -5. -18. PA PA-18:/18:1 71.1 81. -6. -8. -6. -5. -18. PA PA-16:/: 73.1 311.3-6. -8. -6. -5. -18. PA PA-18:/18: 73.1 83.3-6. -8. -6. -5. -18. PA PA-18:/: 73.1 33. -6. -8. -6. -5. -18. PA PA-18:1/:3 73.1 83.3-6. -8. -6. -5. -18. PA PA-3:3 61. 153. -6. -8. -6. -5. -18. PA PA-3: 63. 153. -6. -8. -6. -5. -18. PA PA-3:1 65. 153. -6. -8. -6. -5. -18. PA PA-3: 67. 153. -6. -8. -6. -5. -18. PA PA-3: 671.1 153. -6. -8. -6. -5. -18. PA PA-3:1 673.1 153. -6. -8. -6. -5. -18. PA PA-3: 675.1 153. -6. -8. -6. -5. -18. PA PA-36: 699.1 153. -6. -8. -6. -5. -18. PA PA-36:1 71.1 153. -6. -8. -6. -5. -18. PA PA-36: 73.1 153. -6. -8. -6. -5. -18. PA PA-38: 73.1 153. -6. -8. -6. -5. -18. FFA 1: 199.1 181.1 -. -5. -6. -5. -18. FFA 1: 7.1 183.1-35. -. -6. -5. -18. FFA 1:1 5.1 7.1-5. -. -6. -5. -18. FFA 15: 1.1 197.1-5. -. -6. -5. -18. FFA 16: 55.1 37.1-5. -. -6. -5. -18. FFA 16:1 53.1 9. -35. -. -6. -5. -18. FFA 17: 69. 51. -5. -8. -6. -5. -18. FFA 18:3 77. 33. -5. -. -6. -5. -18. FFA 18:3 77. 59.1-5. -. -6. -5. -18. FFA 18: 79. 61. -55. -. -6. -5. -18. FFA 18:1 81. 63. -. -5. -6. -5. -18. FFA 18: 83.3 65.1-53. -8. -6. -5. -18. FFA :5 31. 57. -5. -18. -6. -5. -18. FFA : 33. 85. -5. -. -6. -5. -18.
FFA :3 35. 1. -65. -5. -6. -5. -18. FFA :3 35. 87. -55. -3. -6. -5. -18. FFA : 37.1 1.3-65. -5. -6. -5. -18. FFA : 39. 1.1 -. -5. -6. -5. -18. FFA : 311.3 18. -65. -5. -6. -5. -18. FFA :6 37. 83. -7. -17. -6. -5. -18. FFA :5 39. 85. -5. -17. -6. -5. -18. FFA : 331. 87. -55. -18. -6. -5. -18. FFA : 335. 69. -65. -3. -6. -5. -18. FFA :1 337. 68.7-5. -5. -6. -5. -18. FFA : 339. 18.9-6. -3. -6. -5. -18. FFA ; 365.3 55.1-5. -35. -6. -5. -18. FFA :1 367.3 39. -5. -35. -6. -5. -18. DP;Declustering potential CE; Collision energy EP; Entrance potential CXP; Collision cell exit potential CEP; Collision cell entrance potential
A B Klotho +/+ Klotho -/- C Klotho +/+ Klotho -/- 5 1 1 35 15 1 3 8 5 1 6 15 1 5 5 Serum Phosphorus (mg/dl) FGF3 (pg/ml) Calcified lesion (µm ) Klotho +/+ Klotho -/- Supplemental Figure 1
Serum creatinine (µg/dl) Calcified lesion (%) Serum stearate (mg/ml) A B C D 3 1 15 1 5 Sham CKD Serum phosphorus (mg/dl) Aortic Calcium (µg/mg) 1 1 1 8 6 18 16 1 1 1 8 6 1. Scd mrna level (vs. sham) 7 6 5 3 1 E F G H 1. 1.8.6.. Scd1 Scd I J K..15.1.5. Kidney function egfr Normal 8 ± 11.5 CKD 3 ± 6.5 Serum oleate (mg/ml)..15.1.5. FGF3 (pg/ml) Normal 8 ± 11.5 CKD 3 ± 6.5 Desaturation index SCD activity (nmol/min/mg protein) 3 1 Sham CKD..3..1 Normal 8 ± 11.5 CKD 3 ± 6.5
G Supplemental Figure 3 Scd mrna (relative mrna) 1.8 1.6 1. 1. 1.8.6.. Scd1 shrna Scd shrna p-eifα ATF CHOP GAPDH 6 5 3 1 A E Scd1 mrna Scd mrna - - + + + - - + empty shrna Scd1 shrna empty shrna Scd shrna Tunica mycin Thapsig argin Calcium (mg/mg protein) 9 8 7 6 5 3 1 Scd1 shrna Scd shrna H Relative Atf6 mrna level K Calcium (µg/mg) 1. 1..8.6.. 3 5 15 1 5 B C Empty shrnas - - + + Scd1 shrna empty shrna empty shrna Scd shrna empty shrna Atf6 shrna I Relative Ire1 mrna level L + - - + ALP activity (unit/mg protein).5 3.5 3.5 1.5 1.5 Scd1 shrna Scd shrna Relative mrna level - - + + + - Empty/Empty shrna Scd1/Scd shrna Scd1/Empty shrna Scd/Empty shrna 7 6 5 3 1 Chop sxbp-1 empty shrna empty shrna empty shrna Atf6 shrna Ire1 shrna Perk shrna 5 35 3 3 5 15 1 1 5 Calcium (µg/mg) Scd1/ shrnas 1. 1..8.6.. F D empty shrna Ire1 shrna J Relative Perk mrna level M Calcium (µg/mg) 1. 1..8.6.. empty shrna Perk shrna - +
% remaining mrna level 1 1 8 6 Empty shrna Acat1 shrna Acat shrna Agpat1 shrna Agpat shrna Agpat3 shrna Agpat shrna Agpat5 shrna Dgat1 shrna Dgat shrna Gpat1 shrna Gpat3 shrna Gpat shrna Lpcat1 shrna Lpcat shrna Lpcat3 shrna Lpcat shrna Lpgat1 shrna Mboat1 shrna Mogat shrna Sptlc1 shrna Supplemental Figure
Ocn mrna level A B C GPAT1 GAPDH GPAT GAPDH 5 3 1 D Calcium (mg/mg) Control shrna Control shrna 6 5 3 1 Gpat1 shrna Gpqt shrna GPAT activity (nmol/min/mg) Control shrna Gpat shrna Gpat1 shrna.6.5..3..1 Calcium (mg/mg) 18: F G H I J ATF CHOP GAPDH 6 Opn mrna level 5 3 1 Gpat shrna E Empty shrna Gpat shrna K ATF 16 6 16 CHOP 16 16 GAPDH 5 15 1 5 PiT1 mrna level 5 3 1 EmptyshRNA EmptyshRNA 1 8 Gpat shrna Pi uptake pmol/mg protein/ min 6 L Atf mrna level 7 6 5 3 1 M Chop mrna level 9 8 7 6 5 3 1 N sxbp-1 mrna level 1 1 1 8 6 Supplemental Figure 5
A B Dox GPAT GAPDH GPAT activity (nmol/min/mg) 1. 1..8.6.. - Dox +Dox C Calcium (µg/mg protein) D Alp mrna level 35 3 3 3 15 1 5 5 3 1 G H I Atf mrna level 5 3 1 Chop mrna level 1 1 1 8 6 18: ATF CHOP GAPDH Dox E PiT1 mrna level F Pi uptake pmol/mg protein/3 min 8 7 6 5 3 1 8 6 1 Supplemental Figure 6
A 5 15 Agpat3 shrna 1 5 8 6 7 6 5 3 1 G 5 Empty shrna ATF CHOP GAPDH 3 1 J Empty shrna 6 ATF CHOP GAPDH Supplemental Figure 7 Agpat5 shrna 8 Empty shrna Agpat5 shrna Agpat3 shrna 1 Empty shrna Agpat3 shrna I H PiT1 mrna level 1 F Ocn mrna level 7 6 5 3 1 E Alp mrna level Ocn mrna level Alp mrna level 3 D Empty shrna PiT1 mrna level Calcium (µg/mg) 3 C B Empty shrna Agpat3 shrna Agpat5 shrna
A mrna expression (pg)/18s(ng) 3.5 3..5. 1.5 1..5 ND Lipin isoform 1 3 E Ocn mrna level H Atf mrna level 7 6 5 3 1 1 8 6 B Chop mrna level F Opn mrna level 1 8 6 16 1 8 Empty shrna K Lipin mrna level 1. 1..8.6.. I C PAP activity (nmol/min/mg) 5 3 1 PiT1 mrna level J sxbp-1 mrna level G 1 1 1 8 6 5 3 1 Lipin shrna D Calcium (mg/mg) 7 6 5 3 1 Empty shrna Lipin shrna 18: ATF CHOP GAPDH Supplemental Figure 8
A B 1 C-18: 1 C-18:1 NL 1.6 vehicle CER PA PE LPA PS PI PC SM LPC 1 C-18:1 incorporation (Fold change) 1. 1. 1.8.6.. PA LPA LPC PI NL PC CER PS PE SM C PA (µg/mg protein) 1. 1.8.6.. sham 5/6nx 1:/18: 16:/16: 16:/18: 16:/: 16:/:1 16:/18: 16:/16:1 16:/18:1 16:1/18: 16:1/18:1 18:/18: 18:/18:1 18:/18: 18:/:3 18:/: 18:1/18:1 18:1/18: D PA content in ER (µg/mg ER protein) 3 5 15 1 5 vehicle 1:/18: 16:/16: 16:/18: 16:/: 16:/:1 16:/18: 16:/16:1 16:/18:1 16:1/18: 16:1/18:1 18:/18: 18:/18:1 18:/18: 18:/:3 18:/: 18:1/18:1 18:1/18: Supplemental Figure 9
A Native Scd Chromosome Floxed Chromosome FLP-FRT Recombination Deleted Chromosome 1 3 5 6 1 3 5 6 PGK- NEO 1 3 5 6 1 5 6 B Scd1/ double lox icle i.p. x 5 Tamoxifen i.p. x 5 Control SMC-Scd1/ double conditional Smmhc-Cre ERT SMC-Scd1/ KO Supplemental Figure 1
Supplemental Figure 1. Serum phosphorous, serum FGF3 and vascular calcification in Klotho -/- mice. A) Phosphorous and B) FGF3 in SMMHC-GFP; Klotho -/- mice and SMMHC- GFP; Klotho +/+ mice (N=8). C) Quantification of calcified lesions in the aortic sinuses of SMMHC-GFP; Klotho -/- mice and SMMHC-GFP; Klotho +/+ mice. The lesions of aortic sinuses were cryosectioned and stained with von Kossa. Supplemental Figure. CKD reduces SCD activity in DBA/J mice and human patients. 8-week-old DBA/J male mice fed a chow diet (N=6) were 5/6 nephrectomized or shamoperated and fed a Western diet (TD11198, Harlan Teklad) for 1 weeks. A) Serum creatinine, B) serum phosphorus and C) FGF3 levels in sham-operated (blue) and 5/6 nephrectomized (red) DBA/J mice. D) Representative picture of aortas with von Kossa staining. E) Quantification of calcified lesions in the aortas of sham-operated and 5/6 nephrectomized DBA/J mice. The aortas were cryosectioned and stained with von Kossa. F) Aortic calcium content in sham-operated and 5/6 nephrectomized DBA/J mice. Aortic calcium level was analyzed using an ash assay coupled with a colorimetric assay. G) Levels of Scd1 and Scd mrna in the aortic medial layers of sham-operated and 5/6 nephrectomized DBA/J mice. H) Aortic SCD activity in sham-operated and 5/6 nephrectomized DBA/J mice. I) Serum stearate, J) serum oleate and K) desaturation index (18:1/18:) in age-matched human patients (N=1 per group) with normal kidney function and stage 3 and CKD was measured by gas chromatography using heptadecanoic acid as an internal standard. Other parameters were previously reported (9). p<.5 and p<.1 (-tailed Student s t-test). Supplemental Figure 3. Scd1 and Scd dual knockdown but not Scd1 and Scd single knockdown induces mineralization, osteogenic differentiation and ER stress of VSMCs. MOVAS-1 cells were treated with adenovirus containing Scd1 shrna and lentivirus containing Scd shrna. For the dual knockdown, Scd stable knockdown MOVAS-1 cells were treated with adenovirus containing Scd1 shrna at a multiplicity of infection (MOI) of. A) Scd1 and Scd mrna levels in Scd1 and Scd dual knockdown cells. Control cells and Scd stable knockdown cells were treated with adenovirus containing SCD1 shrna or empty shrna for hours. Scd1 and Scd mrna levels were analyzed with real-time qpcr. B) Calcium deposit. Cells treated with empty shrnas and Scd1/ shrna were stained with Alizarin red. C) Calcium content in Scd1 and Scd dual knockdown cells. Control cells and SCD stable knockdown cells were treated with adenovirus containing Scd1 shrna or empty shrna for 7 days in the 1
presence of. mm inorganic phosphate. Calcium was extracted with.1n HCl and analyzed with a colorimetric assay. D) ALP activity in Scd1 and Scd dual knockdown cells. Scd stable knockdown cells were treated with adenovirus containing Scd1 shrna or empty shrna for days. E) Immunoblot analysis of ER stress markers. Control cells and Scd stable knockdown cells were treated with adenovirus containing Scd1 shrna or empty shrna for hours. F) mrna levels of ER stress markers. Control cells and Scd stable knockdown cells were treated with adenovirus containing Scd1 shrna or empty shrna for hours. G) ER stress inducers tunicamycin and thapsigargin induce mineralization of VSMCs. VSMCs were treated with.5 g/ml tunicamycin and.5 M thapsigargin for 7 days. Calcium was analyzed with a colorimetric assay. H-J) Atf6, Ire1 and Perk shrnas efficiently reduced their target genes. K-M) Atf6-, Ire1- and Perk-knockdown attenuated SCD inhibition-induced mineralization of VSMCs. Atf6, Ire1 and Perk mrna levels were analyzed with real-time qpcr. Cells were treated with vehicle () or SCD inhibitor () for 7 days in the. mm inorganic phosphate. p<.5 and p<.1 vs. MOVAS cells treated with lentivirus containing empty shrna and adenovirus containing empty shrna or vehicle () ( tailed Student s t-test). Supplemental Figure. Acyltransferase-knockdown stable VSMCs. ShRNA-mediated knockdown of acyltransferases in VSMCs. MOVAS-1 cells were infected with lentiviruses containing acyltransferase shrnas for 8 hours and selected with 5 g/ml puromycin to generate each acyltransferase stable knockdown VSMC. Levels of acyltransferase mrnas were determined by real-time qpcr. Each shrna reduced its targeted acyltransferase by over 75%. N=3. Supplemental Figure 5. Gpat but not Gpat1 knockdown inhibits SFA inducedmineralization, osteogenic differentiation and ER stress in VSMCs. A) Immunoblot analysis of GPAT1 and GPAT in Gpat knockdown cells. MOVAS-1 cells were treated with Gpat1 and Gpat shrna. A single colony was isolated from MOVAS-1 cells infected with each lentivirus in the presence of puromycin. B) GPAT activity in Gpat knockdown cells. Total GPAT activity was measured in total cell lysates of Gpat1 and Gpat knockdown MOVAS-1 cells. C) Stearateinduced mineralization in Gpat1 and Gpat knockdown cells. Gpat1 and Gpat knockdown cells were treated with M stearate (18:) for 7 days. D and E) -induced mineralization in Gpat1 and Gpat knockdown cells. Gpat1 and Gpat knockdown cells were treated with either vehicle or 3 nm for 7 days. Cells were stained with Alizarin red. F) Ocn, G) Opn and H)
PiT1 mrna levels in Gpat1 and Gpat knockdown cells treated with. Gpat1 and Gpat knockdown cells were treated with 3 nm for hours. I) Phosphate uptake of Gpat knockdown cells treated with for 16 hours. J) Immunoblot analysis of stearate-induced ER stress markers in Gpat knockdown cells. Gpat knockdown cells were treated with for hours. K) Immunoblot analysis of -induced ER stress markers in Gpat knockdown cells. Gpat knockdown cells were treated with 3 nm for hours. L-N) mrna levels of ER stress markers such as Atf, Chop and sxbp-1 in Gpat1 and Gpat knockdown cells treated with. Gpat1 and Gpat knockdown cells were treated with 3 nm for hours. White colmun; empty shrna, Red column; Gpat shrna, Black column; Gpat1 shrna p<.1 (one-way ANOVA). Supplemental Figure 6. Human GPAT overexpression augments SFA inducedmineralization, osteogenic differentiation and ER stress in VSMCs. A) Immunoblot analysis of GPAT. MOVAS-1 cells overexpressing human GPAT were generated using a Tet-ON system. MOVAS-1 cells were treated with lentivirus containing tetr and human GPAT. A single colony was isolated from MOVAS-1 cells infected with lentivirus in the presence of hygromycin and zeocin. MOVAS-1 cells containing GPAT were treated with either vehicle or 1 mg/ml doxycycline (Dox) for hours. B) GPAT activity. MOVAS-1 cells containing human Gpat were treated with either vehicle or 1 mg/ml Dox for hours. Total GPAT activity was measured in the total cell lysates of GPAT overexpressed MOVAS-1 cells. C) Stearate and -induced mineralization in GPAT overexpressed cells. MOVAS-1 cells containing GPAT were treated with either vehicle or 1 mg/ml Dox for hours and then treated with M 18: and 3nM for 7 days. D) Alp and E) PiT1 mrna levels in Gpat overexpressed cells. F) Phosphate uptake in GPAT overexpressed cells. MOVAS-1 cells containing Gpat were treated with 1 mg/ml Dox for hours and then treated with 3nM for hours. G) Atf and H) Chop mrna levels in GPAT overexpressed cells. I) Immunoblot analysis of ER stress markers. MOVAS-1 cells containing GPAT were treated with 1 mg/ml Dox for hours and then treated with 3nM for hours. White column; without Dox treatment, Black column; with Dox treatment, p<.1 (one-way ANOVA). Supplemental Figure 7. Agpat3 and Agpat5 knockdowns inhibit SFA-induced mineralization, osteogenic differentiation and ER stress in VSMCs. A) SCD inhibition induced-mineralization in Agpat3 and Agpat5 knockdown cells. Agpat3 and Agpat5 knockdown 3