Supplementary Figure 1
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1 NSC N2d N5d TNFRSFa TNFRSFb RIPK RelA Madd I-TRAF/TANK GAPDH γ-actin Supplementary Figure
2 A 2 hrs Day : Cell seeding Day 9: +ng/ml TNF-α Day 9: Fixation B Merge Nestin PH-3 Hoechst Acute TNF-α Control C % positive cells Control Nestin TNF-α acute D % positive cells Control * PH3 TNF-α acute Supplementary Figure 2
3 35 Apoptosis in NSC/Diff 3 % double-positive cells Nestin / c-casp 3 TuJ / c-casp 3 GFAP / c-casp 3 Control Chronic TNF-α Supplementary Figure 3
4 Merge Hoechst EdU Dcx Cell Profiler Output Cell Profiler Input Supplementary Figure 4
5 A Day : Surgery 6-OHDA or sham Day 5: Start daily IP Minocycline or NaCl Day 2: 2x EdU IP Day 24: Perfusion LV RMS Striatum SN B Hoechst TH TH Ipsilateral Contralateral IF IF Supplementary Figure 5
6 5 Sham Sham + Minocycline m] m] Distance to LV [µ Distance to LV [µ Distance to LV Distance to LV Disatnce to LV Distance to LV Distance to LV to LV to LV Distance Distance Supplementary Figure 6a
7 6 OHDA 6 OHDA+ Minocycline 25 2 ] Distance to LV [µm # Cell # Cell Distance to LV # Cell Disatnce to LV to LV [mm] Distnce Supplementary Figure 6b
8 A Day : Surgery 6-OHDA Day 5: Start daily IP NS-398 or control Day 2: 2x EdU IP Day 24: Perfusion B merge 6-OHDA Hoechst EdU 6-OHDA + NS-398 C Fold increase EdU+ / total cells (6-OHDA=) *.2 6-OHDA 6-OHDA + NS-398 EdU e D 6-OHDA Hoechst Dcx E Migration: Dcx+ cells per distance category (6-OHDA = ) * 6-OHDA + NS-398-5μm 6-OHDA >5μm 6-OHDA + NS-398 Supplementary Figure 7
9 6 OHDA Distance to LV [µ µm] Distance to LV Dista nce to LV Dista nce to LV LV Distance to LV stance to LV Distance to Di Distance to LV [µ µm] Supplementary Figure 8a
10 OHDA + NS Distance to LV [ µm] Distance to LV Distance to LV Distance to LV Distance to LV Distance to LV Distance to LV Distance to LV Supplementary Figure 8b
11 A Day : Surgery 6-OHDA or sham Day 5: Start daily IP Minocycline or control Day 2: 2x EdU IP Day 24: Perfusion B sham 6-OHDA + Minocycline C merge Hoechst EdU Fold increase EdU+ / total cells (sham=) * sham EdU 6-OHDA + Minocycline Supplementary Figure 9
12 Control Mouse # Edu+ Dcx+ EdU+/Dcx+ %EdU+ %Dcx + %EdU+/Dcx+ Edu+ Dcx+ EdU+/Dcx+ % EdU+ % Dcx + %EdU+/Dcx Sum / Mean Table S SVZ counts of EdU +, Dcx +, EdU + /Dcx + -doublepositive cells after TNF-α or control infusion TNF α Control Mouse # Edu+ Dcx+ EdU+/Dcx+ Cell % EdU+ % Dcx + %EdU+/Dcx+ Edu+ Dcx+ EdU+/Dcx+ % EdU+ % Dcx + %EdU+/Dcx Sum / Mean Table S2 prms counts of EdU +, Dcx +, EdU + /Dcx + -doublepositive cells after TNF-α or control infusion TNF α Sham Sham + Minocycline 6 OHDA 6 OHDA + Minocycline Mouse # EdU+ % EdU+ EdU+ % EdU+ EdU+ % EdU+ EdU+ % EdU Sum/Mean Table S3 prms counts of EdU + cells after sham or 6-OHDA injection and control or minocycline treatment, respectively
13 total per animal per animal 5 µm 5 5 µm >5 µm Treatment Mouse # Dcx+ Mean Dist. Median Dist. Dcx+ cells % of Dcx cells Dcx+ % of Dcx cells Dcx+ % of Dcx cells Sham Sum / Mean Sham + Minocyclin Sum / Mean OHDA Sum / Mean OHDA + Minocyclin Sum / Mean Table S4 Intrastriatal immigration distances of Dcx+ cells from LV-border in Sham or 6-OHDA-lesioned and control or Minocycline treated animals 6 OHDA control 6 OHDA + NS 398 Mouse # Edu+ % EdU+ Edu+ % EdU Sum / Mean Table S5 prms counts of EdU + after 6-OHDA injection and control or NS-398 treatment, respectively
14 total per animal per animal <5µm >5µm Mouse # Dcx+ Mean Dist. Median Dist. Dcx+ cells % of Dcx cells Dcx+ cells % of Dcx cells OHDA Sum / Mean OHDA + NS Sum / Mean Table S6 Intrastriatal immigration distance of Dcx+ cells from LV-border in 6-OHDA-lesioned and control or NS- 398 treated animals, respectively Sham 6 OHDA + Minocycline Mouse # Edu+ % EdU+ Edu+ % EdU Sum / Mean Table S7 Neuroinflammatory effects on Neurogenesis in aged mice. prms counts of EdU + after sham or 6-OHDA injection and control or Minocycline treatment, respectively 6 OHDA control 6 OHDA + Minocycline longterm Mouse # counts Measuretime [Min] counts per min Mouse # counts Measuretime [Min] counts per min M M M M M M M M Sum / Mean Table S8 Results of behavioral experiments on Rotameter after 5mg/kg injection of amphetamine in 6-OHDA lesioned mice with either longterm control or Minocycline treatment (recorded by harness measurement; 4 datapoints per turn)
15 6 OHDA control Dorsal Striatum Corpus Callosum Mouse # Section EdU + cells Mean EdU + cells EdU + ;GST π + cells % (EdU+;GST π+)/edu+ Mean % (EdU+;GST π+)/edu+ EdU + cells GST π + cells % GST π + cells N/A N/A N/A Sum Mean OHDA + Minocycline longterm Dorsal Striatum Corpus Callosum Mouse # Section EdU + cells Mean EdU + cells EdU + ;GST π + cells % (EdU+;GST π+)/edu+ Mean % (EdU+;GST π+)/edu+ EdU + cells GST π + cells % GST π + cells N/A 4 N/A N/A 7. N/A N/A N/A N/A N/A N/A Sum Mean Table S9 EdU and GST- π prevalence in the striatum and corpus callosum of 6-OHDA lesioned mice with either longterm control or Minocycline treatment
16 Aditional Files Additional File Title: Supplementary Figures and Tables Description: this file includes supplementary figures and tables
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