Fig. 3

UBE2T facilitates CHK1 activation, release form chromatin to cytosol for turnover upon IR in HCC cells. a Cell cycle distribution was detected in MHCC-97H cells overexpressing UBE2T at indicated timepoints after IR (4 Gy). The number above the bar indicated the average percentage numbers of cells arrested in the G2 phase from triplicate experiments. b MHCC-97H cells with UBE2T knockdown was treated and analyzed as panel a. c Immunoblotting analysis of total lysates from IR (4 Gy) treated UBE2T overexpressing and control MHCC-97H cells for the indicated proteins. d Total protein lysates were extracted from IR (4 Gy) treated UBE2T silencing MHCC-97H cells and control cells for immunoblotting of the indicated proteins. e The cytosol and chromatin protein fractions of IR (4 Gy) treated UBE2T overexpressing MHCC-97H cells were isolated and analyzed. f The cytosol and chromatin protein fractions of IR (4 Gy) treated UBE2T silencing MHCC-97H cells were isolated and analyzed. g Immunofluorescence staining was used to determine the location of CHK1 in UBE2T overexpressing MHCC-97H cells at indicated timepoints after IR (4 Gy). Scale bar: 20 μM. h Immunofluorescent staining of CHK1 (red) in UBE2T silencing cells at indicated timepoints after IR (4 Gy). i UBE2T-overexpressing MHCC-97H cells were treated with IR (4 Gy) and 160 mM CHX for the indicated timepoints and immunoblotted with CHK1. j UBE2T-silencing MHCC-97H cells were treated with IR (4 Gy) and 160 mM CHX for the indicated timepoints and immunoblotted with CHK1. Scale bar: 20 μM. Data represent the mean ± SD. In (a) and (b), ns, not significant, *P < 0.05 and **P < 0.01, by one-way ANOVA