Fig. 5

Upregulation of Fascin-1 phosphorylation partially restores microglial phagocytosis in primary microglia in vitro. (A) Representative immunocytochemistry images of p-Fascin-1 (red), Fascin-1 (green) and primary microglia (phalloidin staining, red) incubated with Dio-myelin (1 mg/ml, green) at 0 or 5 d in primary microglia treated with or without TPA (10 ng/ml, an agonist of PKCγ). (B) Western blot analysis of Fascin-1, p-Fascin-1, PKCγ and p-PKCγ in primary microglia incubated with myelin (1 mg/ml) at 0 or 5 d treated with or without TPA (10 ng/ml). (C) Quantification of MFI for p-Fascin-1, Fascin-1 at 0 or 5 d in primary microglia treated with or without TPA (10 ng/ml) in (A). (D) Quantification of p-Fascin-1/ Fascin-1 in (A). (E) Quantification of intensity of Dio-Myelin staining/cell in (A). (F) Quantification of Fascin-1, p-Fascin-1, PKCγ and p-PKCγ in primary microglia incubated with myelin (1 mg/ml) at 0 or 5 d treated with or without TPA (10 ng/ml) (relative to GAPDH expression) in (B). (G) Quantification of p-Fascin-1/Fascin-1 in (B). (H) Quantification of p-PKCγ/PKCγ in (B). Scale bar: 20 μm (A). Data are presented as means ± SD. n = 3 independent cultures in (B-H). ^*p < 0.05, ^**p < 0.01, ^***p < 0.001 by oneway ANOVA followed by Tukey’s post hoc test