Fig. 7

Mas1 could be the upstream receptor to restore partial microglial phagocytosis in vitro (A) Representative immunofluorescence images of Mas1 (red) at 0 or 5 d in primary microglia incubated with myelin (1 mg/ml) treated with captopril (22 µg/ml, a Mas1 receptor agonist) or BIM (25 µg/ml, a inhibitor of PKCγ). (B) Representative immunofluorescence images of Fascin-1 (red), p-Fascin-1 (green) and primary microglia (phalloidin staining, red) incubated with Dio-myelin (1 mg/ml, green) at 0 or 5d in primary microglia incubated with myelin (1 mg/ml) treated with captopril (22 µg/ml, a Mas1 receptor agonist) or BIM (25 µg/ml, a inhibitor of PKCγ). (C) Quantification of p-Fascin-1/Fascin-1 in (B). (D) Quantification of intensity of Dio-Myelin staining/cell at 0 or 5 d in primary microglia with captopril or BIM treatment. (E) Western blot analysis of Mas1, PKCγ, p-PKCγ, Fascin-1 and p-Fascin-1 at 0 or 5 d in primary microglia with captopril or BIM treatment. (F) Quantification of MFI for Mas1, PKCγ, p-PKCγ, Fascin-1 and p-Fascin-1 at 0 or 5 d in primary microglia with captopril or BIM treatment in (A) and (B). (G) Quantification of Mas1, PKCγ, p-PKCγ, Fascin-1 and p-Fascin-1 at 0 or 5 d in primary microglia with captopril or BIM treatment (relative to GAPDH expression) in (E). (H) Quantification of p-PKCγ/PKCγ in (E). (I) Quantification of p-Fascin-1/Fascin-1 in (E). Scale bar: 20 μm (A) and (B). Data are presented as means ± SD. n = 3 independent cultures in (C), (D), (F), and (G-I). ^*p < 0.05, ^**p < 0.01, ^***p < 0.001 by oneway ANOVA followed by Tukey’s post hoc test