Figure 3.

CWF19L2 is essential for spermatogenesis and male fertility. A) The schematic showing the insertion of loxp sites to flank exon 6 of the mouse Cwf19l2 gene and deletion of Cwf19l2 in germ cells using Stra8‐Cre. B) PCR‐based genotype of the PD5 mouse tails is shown. C) QPCR analysis of knockout efficiency of Cwf19l2 in testes of adult Cwf19l2‐SKO and control mice. Data are presented as mean ± SD, n = 3, ****P < 0.0001 by two‐tailed Student’ s t‐test. D) Immunoblotting analysis of the knockout efficiency of CWF19L2 in the testes of adult Cwf19l2‐SKO and control mice. ACTIN served as the loading control. E) Immunofluorescence staining analysis of the knockout efficiency of CWF19L2 (green) in testis sections of adult Cwf19l2‐SKO and control mice. DNA was stained with DAPI. Scale bars = 30 µm. F) Fertility tests of adult Cwf19l2‐SKO and control mice showed the cumulative number of pups per mouse. Data are presented as mean ± SD, n = 3, ****P < 0.0001 by two‐tailed Student’ s t‐test. G) Gross morphology of the testes of adult Cwf19l2‐SKO and control mice. Scale bars = 2 mm. H) The ratio of testis weight to body weight in adult Cwf19l2‐SKO and control mice. Data are presented as the mean ± SD, n = 4, ****P < 0.0001 by two‐tailed Student’ s t‐test. I) Hematoxylin staining of testes (top panel, Scale bars = 20 µm) and epididymides (ep) (bottom panel, Scale bars = 50 µm) of adult Cwf19l2‐SKO and control mice. Arrows, apoptotic spermatocytes; asterisks, empty seminiferous tubules. J) Co‐immunofluorescence staining of GCNA1 (green) with SOX9 (red) in testis sections of PD10 Cwf19l2‐SKO and control mice. DNA was stained with DAPI. Scale bars = 5 µm. K) The quantification of GCNA1‐positive cells per tubule in J). Data are presented as the mean ± SD, ****P < 0.0001 by two‐tailed Student’ s t‐test. L) The quantification of SOX9‐positive cells per tubule in (). Data are presented as the mean ± SD, ns: not significant by two‐tailed Student’ s t‐test.