Role of the Rac 1/NADPH oxidase/ROS pathway in salusin-β-induced tubular cell injury. HK-2 cells were pretreated with ROS scavenger NAC (1 mM), NADPH oxidase inhibitor Apo (100 μM), and Rac-1 inhibitor EHT1864 (1 μM) for 30 min, and then transfected with lentivirus expressing salusin-β (MOI = 100) for 48 h. (A) Representative blots showing the protein level of Bcl-2, Bax, cleaved-caspase-3 (C-caspase-3), cleaved-PARP (C-PARP). (B) Quantitative analysis of the protein level of Bcl-2, Bax, cleaved-caspase-3 (C-caspase-3), cleaved-PARP (C-PARP). (C) Cell apoptosis determined with TUNEL assay. Blue fluorescence (Hoechst 33342) shows cell nuclei and green fluorescence (TUNEL) stands for apoptotic cells. (D) Representative blots and quantitative analysis of γH2AX, total and phosphorylated p53 at 48 h after salusin-β overexpression. Scale bar = 50 μm. Values are mean ± SE. *P < 0.05 vs. Control, †P < 0.05 vs. Vehicle (Veh). n = 6 for each group. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)