Fig. 2

RND1 overexpression induced the ferroptosis of GBM in vitro. A Effects of RND1 overexpression on U87 cell growth according to the inverted microscope observation and colony formation assay. Ratio of inactive U87 cells under the inverted microscope and cell growth from colony formation assay were quantified. B The efficacy of Flag-RND1 plasmid was assessed by western blot analysis. C According to the cell viability assay, Fer-1 (10 µM) rescued the growth inhibition of U87 induced by RND1 overexpression after 48 h, while Z-VAD (20 µM) and Nec (20 µM) did not. Z-VAD, (Z-VAD (OMe)-FMK); Fer-1, Ferrostatin-1; Nec, Necrosulfonamide. D The cell viability assay showed that the duration of Fer-1 (10 µM) for 48 h or 72 h significantly inhibited the effect of RND1 overexpression in U87. E U87 cells were transfected with Flag or Flag-RND1 plasmids and treated with Fer-1 (10 μM). Western blot assays were used to measure the RND1, p53 and SLC7A11 protein levels. F–H The lipid ROS assay, glutathione assay and peroxidation assay also revealed a regulation of RND1 on the lipid ROS, GSH and MDA level of U87 cells, which was reversed by Fer-1 (10 µM, 48 h). ROS, reactive oxygen species; MDA, malondialdehyde; GSH, glutathione. I Transfected U87 cells were prepared for transmission electron microscopy observation. *P < 0.05; **P < 0.01; ***P < 0.001; ns, no significance