Figure 4
From: Splicing controls the ubiquitin response during DNA double-strand break repair
When splicing-related factors are depleted, overexpression of RNF8 restores repair of DNA double-strand breaks. (a) Western blotting of SF3B1, RBMX, PRPF8 and RNF8 in U2OS cells treated for 48 h with the siRNAs indicated. β-Actin was used as a loading control. (b) U2OS cells were transfected with the siRNAs indicated for 48 h, exposed to IR (6 Gy), and 1 h later, immunostained for γH2AX and conjugated ubiquitin (with the FK2 antibody). (c) U2OS cells were treated with the siRNAs indicated for 40 h, followed by transfection with either GFP-RNF8 or GFP-Empty plasmid for 7 h; exposure to IR (6 Gy) and fixation 1 h later and immunostaining for conjugated ubiquitin. The white numbers indicate the percentage of 100 transfected cells, i.e, green cells whose nuclei contained >10 IR-induced foci. Means±S.D. are shown, n=3. *P<0.05, as determined by a non-paired two-tailed Student’s t-test. (d) Schematic model of how splicing regulates repair of DNA double-strand breaks
