Fig. 5

Properdin binds to and activates the Mincle receptor on microglia. a Description of Mincle receptor-mediated downstream signaling pathways. b Expression of Clec4e in rmProperdin-treated microglia versus control microglia was evaluated by qPCR. n = 3 to 4 per group, ***P < 0.001, unpaired Student’s t test. c Mincle receptor expression was assessed by Western blotting. n = 3 per group, **P < 0.01 versus the control group, one-way ANOVA with Bonferroni post hoc test. d Representative images of the Duolink PLA assay showing the PLA signals elicited by the interaction of rmProperdin and the Mincle receptor on primary microglia. e After immunoprecipitation with anti-Mincle, lysates of primary microglia treated with or without 8 µg/ml rmProperdin were immunoblotted with anti-properdin and anti-Mincle antibodies. Mouse IgG served as a control. f, g Western blotting analysis of p-Syk, p-p65, and C/EBPβ expression in rmProperdin-treated microglia versus control microglia. Quantification of the p-Syk/Syk ratio, p-p65/p65 ratio and C/EBPβ expression was normalized to the expression of the housekeeping gene GAPDH. n = 3 per group, *P < 0.05, **P < 0.01 versus the control group, one-way ANOVA with Bonferroni post hoc test