Inhibition of Ezh2 in Stat5b-CA-expressing BMDCs disturbs their tolerogenic signature and their capacity to protect against diabetes in NOD mice. (A,B) Stat5b-CA.BMDCs were pre-incubated for 1 h with the Ezh2 inhibitor GSK343 (3 µM) or vehicle (0.1% DMSO), then cultured for 48 h in the absence or presence of GM-CSF (50 ng/mL) and analyzed for (A) the expression of CD80, CD86, PD-L1, and PD-L2 by FACS and (B) production of cytokines (TGFβ, IL-10, IL-12, and IL-23) by ELISA. (C) BMDCs derived from NOD or NOD.CD11cStat5b-CA mice were cultured in vitro with Ezh2 inhibitor GSK343 (3 µM) or vehicle (0.1% DMSO). DCs were washed and i.v. injected (107 cells/mouse) to 8-week-old female NOD mice (6 mice per group). Recipient NOD mice were followed for diabetes development until 26 weeks (22 weeks post-injection) of age. The asterisks indicate statistically significant differences determined by one-way ANOVA with Tukey’s post-test. p < 0.05 (*). For diabetes development, data were analyzed by Gehan–Breslow–Wilcoxon test. p < 0.01 (**) and p < 0.001 (***).