Fig. 6

Microglia modulates endothelial CXCL10 production in a VDR-dependent manner. A Illustration of the in vitro co-culture and transmigration system. The former contains primary brain endothelial cells (ECs) in the lower chamber and primary microglia isolated from neonatal control and Vdr-cKO mice in the upper chamber. Co-culture media in the lower chamber are used as conditioned media to stimulate T cell migration in the latter transmigration system. B CXCL10 concentrations in the co-cultures containing control and Vdr-cKO microglia (n = 6 per group). C Quantification of immunofluorescence intensity of CXCL10 in co-cultured ECs of indicated groups 48 h after OGD (n = 6 per group). Scale bar, 20 µm. D The counts of T cells migrating towards the lower chamber (n = 6 per group). E Illustration of the co-culture system containing bEnd.3 and BV2 cells. F, G Quantification of CXCL10 expression in bEnd.3 cells cultured alone, co-cultured with control, VDR-deficient, or VDR-overexpressing BV2 cells, as measured by FACS (F) and ELISA analysis (G) (n = 5 per group). H, I Quantification of CXCL10 expression in bEnd.3 cells of indicated groups treated by anti-TNF-α and/or anti-IFN-γ antibodies, as measured by FACS (H) and ELISA analysis (I) (n = 3 per group). Each symbol represents one biological replicate. Data are expressed as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Two-tailed Student’s t test for B–D; for the former four columns in F, G, one-way ANOVA followed by Dunnett's post hoc test was used to compare each column with the second one; for the latter two columns, two-tailed Student’s t test was used. For the former three columns in H, I, one-way ANOVA followed by Dunnett's post hoc test was used to compare each column with the second one; for the latter four columns, one-way ANOVA followed by Tukey's post hoc test was used to compare each column with the others