Journal: NPJ Parkinson's Disease
Article Title: Systemic inflammation accelerates neurodegeneration in a rat model of Parkinson’s disease overexpressing human alpha synuclein
doi: 10.1038/s41531-024-00824-w
Figure Lengend Snippet: a Representative flow cytometry dot plots of isolated cells illustrating the gating strategy to determine profiles based on CD45 low CD11b + for microglia, CD45 high CD11b high macrophages for the central nervous system (CNS)-associated phagocytes, and CD45 high CD11b− cells. b Percentage of CD45 high CD11b high cells in the SNpc (left) and in the striatum (right). Error bars represent ±SEM. ( n = 5 SAL WT, n = 5 LPS WT, n = 6 SAL Snca +/+ , n = 6 LPS Snca +/+ ; Ordinary two-way ANOVA for Genotype vs Treatment; SNpc: Interaction, F (1, 18) = 0.003483; Genotype, F (1, 18) = 79.06, p < 0.001; Treatment, F (1, 18) = 18.48, p = 0.0004; * p < 0.05, **** p < 0.0001, with Bonferroni’s post hoc multiple comparisons test; striatum: Interaction, F (1, 18) = 0.02436; Genotype, F (1, 18) = 52.37, p < 0.0001; Treatment, F (1, 18) = 13.76, p = 0.0016; *** p < 0.001, **** p < 0.0001, with Bonferroni’s post hoc multiple comparisons test). c Percentage of CD45 low CD11b + microglia in the SNpc (top) and in the striatum (bottom). Error bars represent ±SEM. ( n = 5 SAL WT, n = 5 LPS WT, n = 6 SAL Snca +/+ , n = 6 LPS Snca +/+ ; Ordinary two-way ANOVA for Genotype vs Treatment; SNpc: Interaction, F (1, 18) = 0.7841; Genotype, F (1, 18) = 0.7550; Treatment, F (1, 18) = 9.796, p = 0.0058; striatum: Interaction, F (1, 18) = 0.1473). d Mean fluorescence of CD86 both in the SNpc (top) and striatum (bottom). ( n = 5 SAL WT, n = 5 LPS WT, n = 6 SAL Snca +/+ , n = 6 LPS Snca +/+ ; Ordinary two-way ANOVA for Genotype vs Treatment; SNpc: Interaction, F (1, 18) = 2.858; Genotype, F (1, 18) = 79.88, p < 0.0001; Treatment, F (1, 18) = 9.063, p = 0.0075; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, with Bonferroni’s post hoc multiple comparisons test; striatum: Interaction, F (1, 18) = 4.848, p = 0.0410; Genotype, F (1, 18) = 111.9, p < 0.0001; Treatment, F (1, 18) = 25.47, p < 0.0001; ** p < 0.01, *** p < 0.001, ****p < 0.0001, with Bonferroni’s post hoc multiple comparisons test). e Mean fluorescence of MHC-II both in the SNpc (top) and striatum (bottom). ( n = 5 SAL WT, n = 5 LPS WT, n = 6 SAL Snca +/+ , n = 6 LPS Snca +/+ ; Ordinary two-way ANOVA for Genotype vs Treatment; SNpc: Interaction, F (1, 18) = 0.2921; Genotype, F (1, 18) = 56.27, p < 0.0001; Treatment, F (1, 18) = 623.2, p < 0.0001; *** p < 0.001, **** p < 0.0001, with Bonferroni’s post hoc multiple comparisons test; striatum: Interaction, F (1, 18) = 7.674, p = 0.0126; Genotype, F (1, 18) = 24.36, p = 0.0001; Treatment, F (1, 18) = 40.04, p < 0.0001; *** p < 0.001, **** p < 0.0001, with Bonferroni’s post hoc multiple comparisons test).
Article Snippet: For immunophenotyping of substantia nigra and striatum, single cell suspension was stained on the cell surface in different panels with PerCP5.5-conjugated anti-CD45 (1:100, BioLegend Cat# 103132 (also 103131), RRID:AB_893340), APC-Vio770-conjugated anti-CD11b/c (1:100, Miltenyi Biotec Cat# 130-121-054, RRID:AB_2752215), APC-conjugated anti-CD161 (1:100, BioLegend Cat# 205606, RRID:AB_11142680) or CD86 (1:100, Miltenyi Biotec Cat# 130-109-130, RRID:AB_2659403), PE-conjugated anti-CD45RA (1:100, BioLegend Cat# 202307, RRID:AB_314010), PE-Vio770-conjugated anti-CD3 (1:100, Miltenyi Biotec Cat# 130-103-773, RRID:AB_2657103) or anti-MHC-II (1:100, Miltenyi Biotec Cat# 130-107-822, RRID:AB_2652895), and FITC-conjugated anti-granulocytes (1:100 Miltenyi Biotec Cat# 130-108-084, RRID:AB_2651886) or VioBright-conjugated anti-ACSA-2/O4/CD31 (1:100, Miltenyi Biotec) for 15 minutes at 4°C in the dark.
Techniques: Flow Cytometry, Isolation, Fluorescence
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