Fig. 4.

Oxidative mediated apoptosis and ERS in a H/R cell model. HK-2 cells were pretreated with 5 mM N-acetyl-cysteine (NAC) for 1 h, and then subjected to H/R. (A) Brd4 protein levels were detected by western blot analysis at various reoxygenation time, including 2 h, 4 h, and 6 h, and bar graph showing the fold changes of Brd4 relative to sham group from three independent samples. (B–C) ROS and H₂O₂ production were measured and bar graph quantification of ROS accumulation from three independent experiments. (D) Apoptotic rates of normal cells and cells exposed to H/R were detected and bar graphs represent three independent experiments, each performed in triplicates. (E) Western blot analysis for the protein expression of Bax, BCL-2, caspase-3, and protein levels were quantified by densitometry and normalized to the expression of GAPDH from three independent experiments. (F) Western blot analysis for the protein expression of GRP78, CHOP, and protein levels were quantified by densitometry and normalized to the expression of GAPDH from three independent experiments. Values are expressed as the mean ± SEM. *P < 0.05, relative to control group; #P < 0.05, relative to the H/R+DMSO.