Fig. 5

Nrf2 knockdown inhibited autophagy, attenuated antioxidant, anti-inflammatory activities of SFN in LPS-induced primary GMECs. Cells were transfected with si-Nrf2 for 48 h in advance and then treated with SFN (5 μmol/L) for 12 h in the absence or presence of LPS (10 μg/mL) for 6 h. A Western blot analysis of Nrf2 protein (n = 3). Cells were transfected with negative control (NC) and siRNA against Nrf2 (si-Nrf2) for 48 h. One-way ANOVA, Dunnett's post-hoc test. B Immunofluorescent analysis of LC3 puncta (green) and nuclear staining with DAPI (blue) (Bar = 20 μm, n = 3). C and D Analysis of ROS production (Bar = 100 μm). Two-way ANOVA, Tukey's post-hoc test. E–G qRT-PCR analysis of relative mRNA levels of inflammatory factors (n = 3). Two-way ANOVA, Tukey's post-hoc test. H Immunofluorescent analysis of NF-κBp65 and nuclear staining with DAPI (blue) (Bar = 10 μm). Data are presented as the mean ± SEM. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001 and ^****P < 0.0001