Fig. 4

DMI inhibits MG activation and inflammatory cytokine production and enhances Nrf2/HO-1 defense pathway in MG. Primary MG or BV2 cells were left untreated (MED) or pretreated with vehicle or DMI 150 μM for 1 h followed by LPS (100 ng/ml) stimulation. a 24 h after LPS stimulation, primary MG were subjected to CD68 staining to assess MG activation. The representative confocal images of three independent experiments are shown. Scale bar, 20 μm. b 1.5 h after LPS stimulation, primary MG were collected and subjected to RNA extraction followed by Q-PCR analysis for mRNA expression of GM-CSF, IL-23p19, IL-12p35, IL-12p40, IL-1α, and IL-1β. The representative results of three independent experiments are shown. Statistical significance was determined as **p < 0.01 and ***p < 0.001 by one-way ANOVA with post-hoc Bonferroni’s multiple comparison test. c 1.5, 3, and 5 h after LPS stimulation, BV2 cells were collected and subjected to western blot analysis for Nrf2 and HO-1 expression. The representative results of three independent experiments are shown. d 8 h after LPS stimulation, primary MG were subjected to immunofluorescence analysis to determine HO-1 expression. The representative confocal images of four independent experiments are shown. Scale bar, 20 μm