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. 2017 Nov 13;45(22):12862–12876. doi: 10.1093/nar/gkx1102

Figure 4.

Figure 4.

RBM45 participates in DDR and is important for efficient DNA repair. (A) HeLa cells expressing siRNA targeting NC or RBM45 were stained with γH2AX antibody 30 min later after X-ray (1 Gy) irradiation. Cells with more than ten γH2AX foci were counted. P values from unpaired t test were included. Western blot analysis verified the efficiency of siRNAs targeting RBM45 in HeLa cells. NC: negative control. (B) RBM45 depletion confers modest radio-sensitivity in HeLa cells. Cells expressing siNC or siRBM45 were irradiated and colony formation assay was performed. Values are presented as means ± SD, n = 3. siNC: negative control. (C) Detection of DSB repair efficiency mediated by homologous recombination (HR). 293T cells expressing siRNAs for NC, RBM45, or BRCA1 were cotransfected with I-SceI and DR-GFP. 48 h later cells were harvested and analyzed by FACS. siBRCA1 was used as a positive control. siNC: negative control. (D) RBM45 overexpression enhances the HR efficiency. 48 h after HA-RBM45 or HA-vector cotransfection with I-SceI and DR-GFP, cells were harvested and analyzed by FACS. (E) Detection of DSB repair efficiency mediated by NHEJ. 293T cells expressing siRNAs for NC, RBM45, or Ku70, were co-transfected with pCherry and pEGFP-Pem1-Ad2 plasmids. 24 h later cells were harvested and analyzed by FACS. siKu70 was used as a control. siNC: negative control. (F) RBM45 overexpression enhances the NHEJ efficiency. 24 h after HA-RBM45 or HA-vector cotransfection with pCherry and pEGFP-Pem1-Ad2 plasmids, cells were harvested and analyzed by FACS. P values from unpaired t test were included.