Fig. 1 | Nature Communications

Fig. 1

From: The concerted roles of FANCM and Rad52 in the protection of common fragile sites

Fig. 1

FANCM is important for suppressing Flex1-induced mitotic recombination independent of the FA core complex and ID2 complex. a Schematic drawing of the EGFP-based HR-Flex and HR-Luc reporters as previously described10. Luc: luciferase fragment; D-EGFP: donor EGFP. b, c shRNAs for indicated proteins or a control vector (Ctrl) were expressed in U2OS (b) or Hela and MCF7 (c) cells containing the HR-Flex reporter, and assayed for spontaneous mitotic recombination after culturing for 3 or 12 days. The expression of indicated protein was examined by western blot analysis. d Spontaneous recombination was examined in HCT116 wild-type (WT) and HCT116 FANCM KO cells carrying the HR-3B/AT (AT-rich sequences derived from FRA3B) reporter after culturing pre-sorted non-green cells for indicated days. e, g Spontaneous recombination was assayed in U2OS (HR-Flex) cells expressing Flag-FANCM WT or indicated mutants 12 days after endogenous FANCM was silenced by shRNA. The expression of Flag-FANCM WT and indicated mutants is shown by western blot analysis. f The cell lines described in b were infected with I-SceI retroviruses and assayed for HR 5 days later. In all experiments, error bars represent standard deviation (SD) of at least three independent experiments. The P value is indicated as *P < 0.05 and **P < 0.01. n.s. not significant. Western blot analysis was performed using β-actin or Ku70 as a loading control