Figure 2
From: The deubiquitylating enzyme UCHL3 regulates Ku80 retention at sites of DNA damage
UCHL3 promotes cellular survival after IR and DSB repair. (A,B) U2OS cells transfected with the indicated siRNAs were processed for immunoblotting (A) or were subjected to clonogenic survival assay after IR (B) (Mean ± SEM, n = 3). (C,D) U2OS (wild-type: WT) or U2OS UCHL3 KO (KO#1 and KO#2) cells were processed for immunoblotting (C) or were subjected to clonogenic survival assay after IR (D) (Mean ± SEM, n = 3). (E) U2OS cells transfected with the indicated siRNAs were subjected to neutral comet assay. Efficiency of DSB repair is measured as the tail moment ratio between 2 hours after phleomycin removal and immediately after treatment (Mean ± SEM, n = 3). (F) U2OS cells stably expressing FLAG-UCHL3 or FLAG (empty vector: EV) were transfected with the indicated siRNAs and then subjected to neutral comet assay (Mean ± SEM, n = 3). *p < 0.05, **p < 0.01, ***p < 0.001. The tail moments measured immediately after phleomycin treatment, which indicate the amount of generated DSB, are presented in Supplementary Fig. S3. Full-length blots are presented in Supplementary Fig. S9.
