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. 2016 Nov 3;17(11):1835. doi: 10.3390/ijms17111835

Figure 7.

Figure 7

Grx1 and Nrf2 knockdown abolish the cytoprotective effects of Sal B. RPE cells were transfected with scramble or Grx1 siRNA and then incubated with or without 50 μM Sal B for additional 24 h followed by H2O2 treatment for 6 h. (A) Western blot analysis of Grx1 protein levels. Total protein of each sample was subjected to Western blot with Grx1 and GAPDH antibodies. Bottom panel: the relative pixel density of Grx1 over GAPDH. *** p < 0.001 compared with the scramble siRNA control group (n = 3); (B) Grx1 knockdown abolished the cytoprotective effects of Sal B. Cell viability in each group was measured using a WST-8 assay. Values are the mean ± SEM of three experiments, n = 6; *** p < 0.001 compared with the H2O2-treated group; (C) Western blot analysis of Grx2 protein levels. RPE cells were transfected with scramble or Grx2 siRNA and then incubated with or without 50 μM Sal B for additional 24 h followed by H2O2 treatment for 6 h. Total protein of each sample was subjected to Western blot with Grx2 and GAPDH antibodies. Bottom panel: The relative pixel density of Grx2 over GAPDH. *** p < 0.001 compared with the scramble siRNA control group (n = 3); (D) Grx2 knockdown did not change the cytoprotective effects of Sal B. Cell viability in each group was measured using a WST-8 assay. Values are the mean ± SEM of three experiments, n = 6; ** p < 0.01, *** p < 0.001 compared with H2O2-treated group; (E) Nrf2 gene silencing inhibited Sal B-mediated Grx1 induction. RPE cells were transfected with scramble or Nrf2 siRNA and then incubated with or without 50 μM Sal B for additional 24 h followed by H2O2 treatment for 6 h. Total protein of each sample was subjected to western blot with Nrf2, Grx1, and GAPDH antibodies. Bottom panel: the relative pixel density of Nrf2 and Grx1 over GAPDH. ** p < 0.01, *** p < 0.001 compared with the scramble siRNA control group (n = 3); and (F) Nrf2 siRNA abolished the cytoprotective effects of Sal B. Cell viability in each group was measured using a WST-8 assay. Values are the mean ± SEM of three experiments (n = 6), *** p < 0.001 compared with H2O2-treated group.