FIGURE 6.

Zinc inhibits ferroptosis in VSC4.1 treated with H2O2, thereby reducing lipid peroxide levels. (A) The expressions of NRF2, HO‐1, GPX4, and 4HNE were evaluated by Western blotting in VSC4.1 in each group (n =6). (B‐E) Quantification of NRF2, HO‐1, GPX4, and 4HNE expressions (data shown as mean ± SEM, two‐way ANOVA with Tukey's post hoc test, n = 6). (F) Immunofluorescence staining was used to detect the level of GPX4 from each group (n = 6, scale bar = 50 µm). (G) Statistical analysis of immunofluorescence staining for positive expression of GPX4 in VSC4.1 from each group (n = 6, all the data are expressed as means ± SD, two‐way ANOVA followed by Tukey's post hoc test was applied). (H) Confocal microscopy showed the non‐oxidized lipid (red) and oxidized lipid (green) in VSC4.1 that were pretreated with ZnG (90 μmol/L) or not (n = 6, scale bar = 250 µm). (I) Statistical analysis of the fluorescence intensity of oxidized lipid expression in VSC4.1 from each group (n = 6, all the data are expressed as means ± SD, two‐way ANOVA followed by Tukey's post hoc test was applied). * means p < 0.05; ** means p < 0.01; and *** means p < 0.001