Fig. 5

Elp1 deficiency impairs formation of RAD51 foci by regulating RAD51 protein levels and Rad51 translation. A Representative images of RAD51 foci in Ctrl and KO MEFs without irradiation (0 h) or 1, 4 or 24 h post-irradiation. Scale bars, 10 μm. B Quantification of the number of RAD51 foci per cell, with at least 100 cells counted for each independent experiment. C, D Western blotting analysis (C) and quantitative analysis (D) of RAD51 protein levels. β-actin was used as a loading control. E qRT-PCR analysis of Rad51 mRNA in Ctrl and KO MEFs that were either non-radiated (0 h) or exposed to 4 Gy of IR. Data are from three independent experiments. Results are presented as means ± SEM. Student’s t-test: *p < 0.05; **p < 0.01; ***p < 0.001. F Polysomes from Ctrl and KO MEFs without irradiation (0 h) (top panel) or 4 h post-irradiation (4 Gy) (lower panel) were fractionated by sucrose gradient centrifugation for polysome profile analyses. G qRT-PCR analysis of Rad51 and Gapdh mRNAs in polysomal fractions from (F). The translational activity associated with each fraction is indicated as: non-polysome (not translated); LMW—low-molecular-weight polysomes (moderately translated), or HMW—high-molecular-weight polysomes (actively translated)