Fig. 3

DMF suppresses MG-mediated neurotoxicity. MG were stimulated with LPS (1 μg/ml) in the absence or presence of DMF. Microglial CM was collected and applied to cultured hippocampal neurons. Neurons were exposed to microglial CM for 24 h. a The cell viability was assessed by MTT assay. Data presented are from four independent experiments. *p < 0.05, **p < 0.01, by one-way ANOVA with Bonferroni’s post hoc multiple comparison test. b The Caspase-3 activation was examined by immunostaining to detect cell death in neurons. Cell nuclei were stained blue with Hoechst 33342. Representative images are from three independent experiments. MED, cell culture medium. VEH, vehicle control. Scale bar, 20 μm. c The cleaved Caspas-3⁺ cells were quantified and compared with total cells (Hoechst-stained cells). Data presented are percentage of cleaved Caspas-3⁺ cells in each treatment group. Results 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. d Representative images show a shift of JC-1 fluorescence from orange red (mitochondrial membrane polarized) to green (mitochondrial membrane depolarized) in neurons incubated with CM from LPS-primed MG for 6 h. Neurons incubated with the CM generated from DMF-treated MG (100 μM) show decreased JC-1 green fluorescence. MED, cell culture medium. VEH, vehicle control. Scale bar, 40 μm. Results are from three independent experiments