Fig. 2

Deletion of Rsrc1-161aa leads to abnormalities in spermatogenesis. a Expression levels of Rsrc1 (181 bp) and circRsrc1 (166 bp) were measured by RT-PCR in different mouse tissues. The expression levels are normalized to 18S. b Relative expression levels of circRsrc1 in mouse testes at different ages (in weeks) (n = 3). c Relative expression levels of circRsrc1 in different germ cells and somatic cells of mouse testes (n = 3). d Mass spectrometry detection of circRsrc1 translated peptides from the wild-type and different knockout mice. e The litter size of different knockout and wild-type mice are shown. f–h The sperm count, sperm motile, and sperm progressive of different knockout and wild-type mice (n = 6). i H&E staining of testis sections from adult wild-type and Rsrc1^+/+/circRsrc1^−/− mouse. The red asterisk represents the blocked metaphase. Bar, 10 μm. j,k Quantification of the average number of metaphases per seminiferous tubule at stage XII and stage I of wild-type and Rsrc1^+/+/circRsrc1^−/− mice (n = 3). l Flow cytometry analysis of testes in wild-type and Rsrc1^+/+/circRsrc1^−/− mice (n = 13). m The counts of spermatocytes and round sperm and the ratio of round sperm to spermatocytes in the testes of wild-type and Rsrc1^+/+/circRsrc1^−/− mice (n = 3). n TUNEL staining of wild-type and Rsrc1^+/+/circRsrc1^−/− mice at stage I. Bar, 20 μm. o,p Quantification of the proportion of TUNEL positive metaphase per seminiferous tubule in wild-type and Rsrc1^+/+/circRsrc1.^−/− mice at stage XII and stage I. All data are means ± SEM. When performing statistical tests between two groups of data, tested using a two-tailed unpaired t-test. When performing statistical tests between three or more groups of data, tested using one-way ANOVA with Tukey’s post hoc analysis. *P < 0.05; **P < 0.01; ***P < 0.001