Fig. 4
YARS induces necroptotic cell death through reactive oxygen species (ROS) production and mitochondrial dysfunction in breast cancer. a The subcellular components enriched in the Gene Ontology analysis between CR and nCR. b The mitochondrial morphologies of control (pLOC) and YARS-overexpressing cells captured using transmission electron microscopy (TEM) in the presence or absence of H2O2. The scale bar represents 1 μm. c Representative graphs of mitoSOX flow cytometry analysis in the presence or absence of H2O2. d Real-time measurements of the mitochondrial oxygen consumption rates (OCRs) indicating the mitochondrial biogenetic profile (top). Quantification of the basal OCR, ATP-linked OCR, maximal OCR, nonmitochondrial OCR, and proton leak (bottom). e Protein levels of RIPK, p-RIPK, RIPK3, p-RIPK3, MLKL, and p-MLKL were assessed in the presence or absence of z-VAD.fmk in MDA-MB-231 and T47D cells by western blotting (left), and western blot bands were quantified by densitometry using the ImageJ Lab software (right). f Comparative western blot analyses of YARS and SMAC expression in the cytosolic fractions of control and YARS cells (T47D, BT-20, and MDA-MB-231, left) and western blot band densities (right). Three independent western blots were quantified by densitometry using the ImageJ Lab software. GAPDH served as a loading control. Data are expressed as the mean ± SEM of triplicate experiments; *P < 0.05; **P < 0.01; ***P < 0.001