Fig. 7

ACT001 disrupts the feed-forward activation loop between microglia and Th17 cells in vitro. (A-B) Flow cytometry analysis of the inhibitory effect of ACT001 on 2’3’-cGAMP-induced activation of primary microglia (n = 5 per group). (A) Representative flow cytometry results demonstrating the effect of ACT001 on iNOS and CD206 expression in 2’3’-cGAMP-activated primary microglia. (B) Statistical analysis of the frequencies of iNOS⁺ and CD206⁺ microglia in (A). (C) Primary microglia were treated as described in (A) and analyzed for mRNA expression of cytokines related to microglial activation (n = 6 per group). (D-G) In vitro co-culture experiments were conducted to evaluate the activation of microglia and Th17 cells following co-culture. (D) Representative flow cytometry analysis of IL-17 A in the Th17 cells following co-cultured with microglia treated with 2’3’-cGAMP and ACT001. (E) Statistical analysis of the frequencies and absolute numbers of Th17 cells in (D). (F) Normalized mRNA expressions of Il17a and Rorc in the Th17 cells following co-culture as described in (D) (n = 5 per group). (G) Normalized mRNA expressions of Ciita and Il1b in microglia after 8–16 h of co-culture with Th17 cells as described in (D). (n = 5 per group). Statistics were calculated using one-way ANOVA with Tukey’s correction. Error bars represent the mean ± SEM with ns indicating not significant, *p < 0.05, **p < 0.01, and ***p < 0.001