Fig. 5: Effects of Cox5a knockdown on hypoxia-mediated cell death in H9c2 cells.

a Cox5a knockdown via siRNA transfection was validated using qRT-PCR. All qRT-PCR data are shown as the target gene expression level normalized to the Gapdh expression level, and the data are representative of two independent experiments. b Changes in cell viability (left) and cytotoxicity (right) in H9c2 cells with Cox5a knockdown under normoxic and hypoxic conditions were determined by a WST-based cell viability assay and an LDH-based cytotoxicity assay, respectively. Experiments were performed in triplicate, and the data are representative of two independent experiments. c Cell death-related protein levels after Cox5a knockdown were measured using immunoblot analysis. Band intensities were measured as area densities and analyzed using ImageJ software. Relative intensity levels indicate the protein level normalized to the β-actin level. Immunoblot experiments were performed twice independently. Significant differences between groups were determined via ANOVA, with p values indicated as *p < 0.05 and **p < 0.01. N cells under normoxic conditions, H12h cells exposed to hypoxia for 12 h, NC negative control cells, KD Cox5a-knockdown cells.