Fig. 5.

ALZ003 induces lipid peroxidation through decreasing GPX4 expression. A. After 24 h treatment, lipid peroxidation was estimated by staining cells with BODIPY 581/591C11 reagent. Immunofluorescent signal representing peroxidase lipid was photographed by fluorescent microscope under 40X magnification. The scale bar represented 0.1 mm. After treatment for 24 h, GPX4 expression in U87MG (B) and Pt#3 (C) cells was analyzed. Experiments were performed three times independently, and data were expressed as mean ± s.e.m. P-value was determined by Student's t-test compared with the Control group. (*p < 0.05, **p < 0.01). C. Effect of GPX4 on ALZ003-induced lipid peroxidation. (a). Overexpression of Flag-GPX4. *Star represents the non-specific band. After transfection for 24 h, cells were treated with ALZ003 for 24 h. Subsequently cells were subjected to BODIPY staining (b) and H₂O₂-Glo analysis (c) (**p < 0.01, ***p < 0.001; ^#p < 0.05, ^##p < 0.01 compared with the group without treatment and overexpression.) Immunofluorescent signal representing peroxidase lipid was photographed by fluorescent microscope under 40X magnification. The scale bar represented 0.1 mm.