Fig. 4.

Rnaseh1 deletion impairs the DSB repair process in spermatogenic cells. A: Immunofluorescence staining of SYCP3 and γH2AX on WT and Rnaseh1^fl/−;Stra8-Cre spermatocyte spreads. Scale bars: 10 µm. B: The proportion of autosomes DSB repaired and unrepaired ratio in WT and Rnaseh1^fl/−;Stra8-Cre spermatocytes. Error bars, s.e.m. **P < 0.01 (two-tailed Student’s t-test). The experiment was repeated at least 3 times. C-E: Immunofluorescence staining of DMC1 (A), RAD51 (B), and RPA2 (C) on spermatocyte spreads made from WT and Rnaseh1^fl/−;Stra8-Cre mice. Scale bars: 10 µm. F: Western blot of DSB repairing-related proteins in spermatogenic cells isolated from adult WT and Rnaseh1^fl/−;Stra8-Cre mice. L/Z, leptotene/zygotene; P, pachytene. G-I: Signal quantification of DMC1 (E), RAD51 (F), and RPA2 (G) foci number per cell at different meiotic stages. Error bars, s.e.m. ***P < 0.001 (two-tailed Student’s t-test). n.s., non-significant. Above experiments were repeated at least 3 times.