Fig. 3

rmProperdin activates microglia and induces microglia-potentiated neuronal death in vitro. a The mRNA expression of inflammatory factors was measured in primary microglia treated with 2 µg/ml, 4 µg/ml and 8 µg/ml rmProperdin versus control microglia. n = 3 per group, *P < 0.05 versus the control group, **P < 0.01 versus the control group, ***P < 0.001 versus the control group, n.s, no significance (P > 0.05), one-way ANOVA with Bonferroni post hoc test. b FACS analysis of IL-1β, TNF-α and IL-6 expression in primary microglia treated with rmProperdin for 24 h. MFI was quantified. n = 3 per group, **P < 0.01 versus the control group, ***P < 0.001 versus the control group, one-way ANOVA with Bonferroni post hoc test. c Experimental design for the analysis of neuronal viability after treatment with rmProperdin or CM from rmProperdin-treated microglia. d Neuronal viability was assessed by CCK8 assay. n = 6 per group, ***P < 0.001 versus the control group, one-way ANOVA with Bonferroni post hoc test. e Analysis of neuronal death using calcein-AM (green)/PI (red) double staining. Scale bar: 50 µm. f Calcein-AM-positive primary cortical neurons were quantified as percentages of total cells. n = 5 per group, **P < 0.01 versus the control group, ***P < 0.001 versus the control group, one-way ANOVA with Bonferroni post hoc test