Fig. 2

UBE2T promotes radioresistance and improves DDR of HCC after IR exposure. a Colony formation assays were employed in MHCC-97H cells with stable UBE2T overexpression (left, representative images of colony formation; right, survival curve and immunoblotting). b MHCC-97H cells were transfected with a control shRNA lentiviral vector (shControl) or shRNA lentiviral vectors targeting UBE2T (shUBE2T). Lentivirus carrying UBE2T with synonymous mutation which cannot be recognized by shUBE2T was transfected into shUBE2T MHCC-97H cells (shUBE2T + UBE2T). Colony formation assays were conducted. c Transplanted xenografts were established with UBE2T overexpressing (UBE2T) and vector transduced (Control) MHCC-97H cells. Tumor volumes of the xenografts from were measured for 21 days. d Tumor weights of the removed xenografts from panel c were measured. e MHCC-97H cells stably overexpressing UBE2T and control cells were exposed to IR (4 Gy). Cells were prepared for immunofluorescence analysis of γH2AX nuclear foci. The 0 timepoint indicated no IR. Representative images and quantification are shown. Scale bar: 20 μM. f shControl, shUBE2T and shUBE2T + UBE2T MHCC-97H cells were treated and analyzed in the same way as that in panel e. Scale bar: 20 μM. g Cell lysates from the cells with the same treatment as that in panel e were made for immunoblotting of γH2AX and GAPDH. h Cell lysates from the cells with the same treatment as that in panel f were made for immunoblotting of γH2AX and GAPDH. i Comet assay was performed in UBE2T overexpressing MHCC-97H cells at the indicated timepoints after IR treatment. (left, representative images; right, bar charts indicating the average tail moment per cell). j Cells were treated as that in panel f and made for comet assay at the indicated timepoints after IR treatment. Data represent the mean ± SD. In (d), (e), (f), (i) and (j), *P < 0.05 and **P < 0.01, by one-way ANOVA. In (c), *P < 0.05, by two-way ANOVA