Fig. 6.

Brd4 inhibition ameliorated apoptosis and ERS through FoxO4. (A) Representative bands of Western blot analysis for the expression of FoxO4 and bar graphs from three independent experiments. (B–C) ROS production measured by flow cytometry in the indicated groups from three independent experiments and bar graph showing the of quantification ROS, and H₂O₂ production measured by Amplex Red in HK-2 cells, and bar graphs represent three independent experiments, each performed in triplicates. (D) Western Blot analysis for the protein expression of Bax, Bcl-2, Caspase-3 and bar graph quantification as indicated from three independent experiments. (E) Western Blot analysis for the protein expression of GRP78 and CHOP, 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+si-NC. (F) Western Blot analysis for the protein expression of FoxO4, and bar graphs represent three independent experiments. (G) FoxO4 mRNA levels were detected by real-time RT-PCR and bar graph quantification as indicated from three independent experiments, *P < 0.05, relative to H/R; #P < 0.05, relative to H/R+si-NC. (H–L) HK-2 cells were pretreated with JQ1 (10 μm) for 1 h before H/R model established, and HK-2 cells were infected with adenovirus carrying the human FoxO4 for 48 h before being exposed to H/R. (H) Representative Western blot analysis of FoxO4 in the indicated groups. (I) Representative Western blot analysis of Bax, Bcl-2, Caspase-3 in the indicated groups. (J) Western blot analysis for the protein expression of GRP78 and CHOP in the indicated groups. (K–L) ROS and H₂O₂ production were measured. Values are expressed as the mean ± SEM; *P < 0.05 versus H/R+JQ1.