P16 deletion inhibited ROS accumulation and its signaling pathway after ATN. A, B. ROS was detected by flow cytometry. Values are mean ± SEM of six determinations per group. *P < 0.05, **P < 0.01, ***P < 0.001 compared with WT mice at the same time after ATN; ###P < 0.001 compared with control at the same genotyped mice; &P < 0.05, &&&P < 0.001 compared with ATN-24 h group at the same genotyped mice; @@P < 0.01 compared with ATN-48 h at the same genotyped mice. C. Western blots of renal extracts showing AIF, KEAP1 and PGAM5, and β-actin was the loading control. D-F. Protein levels relative to β-actin were assessed by d ensitometric analysis. *P < 0.05, **P < 0.01, ***P < 0.001 compared with control at the same genotyped mice; #P < 0.05, ##P < 0.01 compared with WT+ATN group; &P < 0.05, &&P < 0.01 compared with ATN 24 h at the same genotyped and treatment mice; @P < 0.05, @@P < 0.01 compared with ATN 48 h at the same genotyped and treatment mice.