Fig. 5

INHBA inhibits mitochondrial ferroptosis by upregulating SLC25A10 to activate the mtGSH/GPX4 axis. a TEM of mitochondrial morphology in CRC cells after INHBA silencing. Scale bar: 2 μm (top), 1 μm (bottom). b Detection of mtGSH levels (knockdown of INHBA + restoration of SLC25A10): In the cell model with INHBA knockdown and restored SLC25A10 expression, mitochondria were isolated via a mitochondrial separation kit, and mtGSH levels were detected via ELISA. c Detection of mitochondrial GPX4 protein (knockdown of INHBA + restoration of SLC25A10): In the cell model with INHBA knockdown and restored SLC25A10 expression, mitochondria were isolated via a mitochondrial separation kit, and mitochondrial GPX4 protein was measured by Western blot. d Detection of mtGSH levels (overexpression of INHBA + knockdown of SLC25A10): In the cell model with INHBA overexpression and SLC25A10 knockdown, mitochondria were isolated via a mitochondrial separation kit, and mtGSH levels were detected via ELISA. e Detection of mitochondrial GPX4 protein (overexpression of INHBA + knockdown of SLC25A10): In the cell model with INHBA overexpression and SLC25A10 knockdown, mitochondria were isolated via a mitochondrial separation kit, and mitochondrial GPX4 protein was measured by Western blot. f Detection of lipid peroxidation levels: In human CRC cells with INHBA knockdown and restored SLC25A10 expression, MitoPerOx was used to measure lipid peroxidation within the mitochondrial inner membrane. g Detection of ferrous ion changes: In human CRC cells with INHBA knockdown and restored SLC25A10 expression, changes in ferrous ions in the mitochondria were detected via Mito-FerroGreen. For all the statistical plots, data are means ± SD; n = 3 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001