Fig. 1

DMF suppresses LPS-induced reactive MG. Primary MG were stimulated by LPS (100 ng/ml) in the absence or presence of DMF for various time periods. a Microglial surface markers, CD86, CD80, and CD40, were examined at 18 h after stimulation using FACS analysis. Representative flow plots from five independent experiments are shown. ISO, the isotype control antibody. MED, cell culture medium. b Morphological changes of MG at 18 h after stimulation were examined using Iba1 immunostaining. Arrows indicate activated MG with increased expression of Iba1 and enlarged cell bodies. Cell nuclei were stained blue with Hoechst 33342. Scale bar, 20 μm. c The expressions of DMF-induced Nrf2 target genes were examined at 3 h after stimulation. The mRNA levels of hmox1, nqo1, gclc, and gclm were quantified using qPCR. Data presented are from 3 to 5 independent experiments. *p < 0.05, **p < 0.01, by one-way ANOVA with Bonferroni’s post hoc multiple comparison test. d The Nrf2^+/+ and Nrf2^−/− microglial cells were harvested for western blot analysis to examine Nrf2 and HO-1 proteins at 1.5 and 5 h after stimulation. Representative results are shown from three independent experiments. e The CD86, CD80, and CD40 expressed on the surface of Nrf2^−/− MG were examined at 18 h after stimulation using FACS analysis. Representative flow plots from three independent experiments are shown. f Expressions of inflammatory mediators were examined in Nrf2^+/+ and Nrf2^−/− MG at 3 h after stimulation. The mRNA levels of tnf, il1b, nos2, il23a, and il12b in treated MG were quantified using qPCR. Data presented are from three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, by one-way ANOVA with Bonferroni’s post hoc multiple comparison test.