Fig. 4

MSR1 knockout inhibited pro-inflammatory polarization of macrophages after treatment with myelin debris in vitro. a IF staining of F4/80 (green), iNOS (pro-inflammatory macrophage marker, purple), and CD206 (anti-inflammatory macrophage marker, red) in the lesion sites of the MSR1 WT and KO mice. Bar = 200 μm. b Determination of the pro-inflammatory and anti-inflammatory macrophages by analysis the number of iNOS+ and F4/80+ cells or CD206+ and F4/80+ cells in the lesion sites of the MSR1 WT and KO mice (n = 3 per group, values are the mean ± SD, NS indicates no significance, *p < 0.05, **p < 0.01, two-tailed Student’s t tests). c Quantitative PCR of pro-inflammatory macrophage marker genes (iNOS, TNF-α, and IL-β) and anti-inflammatory macrophage marker genes (CD206, YM1/2, and Arg1) in the lesion sites of the MSR1 WT and KO mice at day 7 post injury (n = 3 per group, values are the mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA). d IF staining of iNOS and CD206 in MSR1 WT and KO macrophages after treatment with myelin debris for 24 h. Scale bar = 10 μm. e Quantitative PCR of pro-inflammatory macrophage marker genes (iNOS, TNF-α, and IL-1β) and anti-inflammatory macrophage marker genes (CD206, YM1/2, and Arg1) in MSR1 WT and KO macrophages before or after treatment with myelin debris (n = 3 per group, values are the mean ± SD, NS indicates no significance, *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA). f–g Flow cytometric analysis of myelin debris-treated macrophages from MSR1 WT or KO mice. The percentages of F4/80+ iNOS+ and F4/80+ CD206+ macrophages in the MSR1 WT and KO groups were calculated (n = 3 per group, values are the mean ± SD, NS indicates no significance, *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA)