Figure 5
CST1 inhibits AF-induced cell death by regulating intracellular ROS. (a) CM-H2DCFDA was used for detection of AF-induced ROS by flow cytometry after 2 h of incubation with 2.5 μM AF. (b) Effect of ROS scavengers NAC (1 mM) and M-TEMPO (100 μM) on AF-induced cell death as measured by WST-1 assay. ROS scavengers were co-treated with 2.5 μM AF for 24 h. (c) Western blot analysis to detect AF-induced cell death after co-incubation of AF and ROS scavengers for 24 h. (d and e) Intracellular ROS levels of CST1-overexpressing LoVo and RKO cells (d) and shCST1 knockdown HT-29 and SW480 cells (e) as measured by flow cytometry after 2 h of incubation with AF. (f) TrxR1 expression and activity in LoVo, RKO, HT-29, and SW480 cells after 12 h of incubation with AF as measured by western blot and TrxR1 assay kit. All data shown are the mean±S.D. of three independent experiments. *P<0.05, #P<0.01 versus control
