Fig. 2

SFN suppressed ROS production, up-regulated expression of antioxidant enzymes and activated Nrf2 signaling in LPS-induced GMECs. Cells were pretreated with different concentrations of SFN (1.25, 2.5 and 5 μmol/L) for 12 h in the absence or presence of LPS (10 μg/mL) for 6 h. A ROS production were detected by DCFH-DA (Bar = 100 μm). B Quantification of ROS levels. One-way ANOVA, Dunnett's post-hoc test (n = 6). C qRT-PCR analysis of Nrf2 mRNA (n = 3). One-way ANOVA, Dunnett's post-hoc test. D and E Western blot analysis of Nrf2 protein (n = 3). One-way ANOVA, Dunnett's post-hoc test. F qRT-PCR analysis of Nrf2-targeted genes HO-1, NQO1, GCLC and GCLM mRNA expression (n = 3). Two-way ANOVA, Dunnett's post-hoc test. G and H Western blot analysis of Nrf2 downstream proteins HO-1, NQO1, GCLC and GCLM (n = 3) levels. Two-way ANOVA, Dunnett's post-hoc test. I Immunofluorescent analysis of Nrf2 and nuclear staining with DAPI (blue) in GMECs pretreated with SFN (5 μmol/L) for 12 h in the absence or presence of LPS (10 μg/mL) for 6 h (Bar = 10 μm). Data are presented as the mean ± SEM. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001 and ^****P < 0.0001