Fig. 5.

JQ1 alleviated oxidative stress, apoptosis and ERS in vitro. (A–B) HK-2 cells were pretreated with or without JQ1 at different doses (0.1, 1, 10 μM) for 1 h, and then were exposed to H/R. (A) Effect of JQ1 with different concentrations on cell viability detected by CCK8 upon H/R stimulation in the indicated groups, and (B) representative bands of Western blot analysis for the expression of Brd4 and bar graph quantification as indicated from three independent experiments, Values are expressed as the mean ± SEM; *P < 0.05, relative to control; **P < 0.05, relative to H/R+DMSO; #P < 0.05, relative to H/R+JQ1 (0.1 μM); & P < 0.05, relative to H/R+JQ1 (1 μM). (C–G) HK-2 cells were transfected with a siRNA against Brd4 or a negative control siRNA (si-NC) for 48 h and were pretreated with or without JQ1 at dose of 10 μM for 1 h before being exposed to H/R. (C) Representative bands of Western blot analysis for the expression of Brd4 and bar graph quantification as indicated from three independent experiments. (D) Apoptotic cells exposed to H/R were detected by flow cytometry, and bar graphs represent three independent experiments, each performed in triplicates. (E) Representative Western blot analysis of Bax, Bcl-2, Caspase-3 and bar graphs from three independent experiments, each performed in triplicates. (F) Representative bands of Western blot analysis for the expression of GRP78 and CHOP, and bar graphs from three independent experiments. (G–H) ROS production was measured by flow cytometry and bar graph showed the quantification of ROS in the indicated groups from three independent experiments, and H₂O₂ production measured by Amplex Red in HK-2 cells, and bar graphs represent three independent experiments, each performed in triplicates. Values are expressed as the mean ± SEM; *P < 0.05, relative to H/R group; #P < 0.05, relative to H/R+si-NC.