Fig. 5: Lysine(K)/Arginine(R) motif in the SYCP2 M1 ___domain is required for R-loop stabilization and TC-HR.
From: Meiotic protein SYCP2 confers resistance to DNA-damaging agents through R-loop-mediated DNA repair

A Schematic of SYCP2 mutants by replacing Lysine(K)/Arginine(R) with Alanine(A) in the M1 ___domain of SYCP2. B Relative HR frequency using CRISPR-based LMNA reporter assay with overexpression of indicated SYCP2 and SYCP2 mutants (n = 3 experiments, Mean +/− SEM). (p = 0.0177). C Schematic and WB of siSYCP2 and siSYCP2#2 were shown. The experiments were repeated three times with similar results. D Fold increase of intensity of SYCP2 (Left) or RAD51 (right) at TA-KR sites in siSYCP2#2 pretreated U2OS cells with or without expression of SYCP2 or its mutant as indicated (n = 6 cells and n = 11 cells, Mean +/− SEM). E–G The numbers of SYCP2 and RAD51 IRIF (n = 200 cells, +/− SEM) (E); relative HR frequency in CRISPR-based LMNA HR assay (F) (n = 3 experiments, Mean +/− SEM), and relative TC-HR frequency (G) (n = 3 experiments, Mean +/− SEM) in indicated cells were shown. Mean frequency of the HR (FACs) compared to the siSYCP2 is shown. (p = 0.0001, p = 0.0002). H siSYCP2#2 pretreated U2OS-TRE cells transfected with TA-KR were light-activated and recovered at 4 hr and stained with anti-S9.6. The frequency of positively stained S9.6 foci at TA-KR was quantified (n = 3 experiments, Mean +/− SEM). Mean frequency of quantity of the positive staining of S9.6 at TA-KR is shown. Statistical analysis was done with the unpaired two-tailed Student-t-test, ****p < 0.0001. Source data are provided as a Source Data file.