Fig. 5: Smad3 knockdown induces ferroptosis in mesenchymal-like bladder cancer cells.

A CCK-8 assay demonstrating the effect of Smad3 knockdown on proliferation. B, C Fe2+ levels in cells visualized by confocal imaging. D Flow cytometry analysis of total intracellular ROS levels after Smad3 knockdown. E LPO levels assessed via an LPO assay kit. F MDA content measured via an MDA assay kit. G, H Representative confocal images of lipid peroxidation. Red indicates non-oxidized lipids, and green indicates oxidized lipids. I, J Transmission electron micrographs revealed mitochondrial alterations following Smad3 knockdown (black arrows). Data are presented as the mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; ns, not significant (p > 0.05).