Fig. 2

Ovol2 is dispensable for spermatogenesis in mice. a Schematic illustration of the targeting strategy for generating a complete inactivation of Ovol2 in mouse testes. Mice containing the floxed allele were crossed with Vasa-Cre mouse lines to generate male germ cell-specific deletion of Ovol2 (Vasa-Cre was activated at embryonic day 15.5, E15.5). P1/2 indicate the position of primers used for detection of floxed and WT alleles. P3/4 indicate the position of primers used for detection of delete allele after Cre recombination. b Representative PCR genotyping results showing that the floxed (lox) and the WT (+) alleles can be detected as larger (223 bp) and a shorter (125 bp) bands, respectively. The last two right lanes are the Vasa-Cre transgene detection. M, marker; NC, negative control. c RT-qPCR analyses of Ovol2 mRNA levels in WT and cKO (Vasa-cre; Ovol2^floxflox) adult testes. Wt1 gene was used as RNA quality control. Data are presented as mean ± SEM, n = 6. **P < 0.01 by student t-test. d Representative Western blot analysis of OVOL2 protein levels in WT and cKO (Vasa-cre; Ovol2^floxflox) adult testes. GAPDH served as a loading control. e Quantification analysis of OVOL2 protein levels in WT and cKO adult testes. Data are presented as mean ± SEM, n = 6. *P < 0.01 by student t-test. f Gross morphology of the testis and the epididymis in WT and cKO adult mice. g Histogram showing the testis weights in WT and cKO adult mice. Data are presented as mean ± SEM, n = 6. h Periodic acid-Schiff (PAS) staining showing the histology of testes from adult WT and cKO mice. Scale bar = 50 μm. i RT-qPCR assays showing elevated Ovol1 mRNA levels in cKO adult testes. Data are presented as mean ± SEM, n = 6. *P < 0.05 by student t-test. j Representative Western blot analysis of OVOL1 protein levels in WT and cKO adult testes. GAPDH served as a loading control. k Quantification analysis of OVOL1 protein levels in WT and cKO adult testes. Data are presented as mean ± SEM, n = 6. *P < 0.05 by student t-test