Figure 6.

RNF4 is required for HR-repair and Rad51 recruitment to DSBs. (a) Graphical representation of the efficiency of Sce–I induced HR in 293 DR-GFP expressing the indicated shRNAs (upper panel). Immunoblot showing RNF4 levels in 293 DR-GFP cells tranfected with the indicated shRNAs; Vinculin serves as loading control (lower panel). (b) Graphical representation of the efficiency of NHEJ DSBs repair, as measured by the proportion of EGFP-positive cells expressing the indicated shRNAs. (c) Graphical representation of the efficiency of NHEJ DSBs repair in Rnf4 +/+ and Rnf4 h/h MEFs. (d) Representative micrographs of MEFs stained with anti-Rad51 antibody and DAPI. Rnf4 +/+ and Rnf4 h/h were untreated (NT) or exposed to 5 Gy of IR and fixed at the indicated time post IR. Bars, 10 μm. (e) Representative micrographs of MEFs stained with anti-Rad51 and anti γ-H2 AX antibodies. Rnf4 +/+ and Rnf4 h/h were untreated (NT) or exposed to 5 Gy of IR and fixed 6h post IR