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. 2023 Dec 1;81(1):1. doi: 10.1007/s00018-023-05025-x

Fig. 1.

Fig. 1

The knockout of Ccdc146 leads to male infertility. A CCDC146 was related to CCDC38 predicted by the STRING database. B CCDC146 interacted with CCDC38. pCSII-MYC-Ccdc146 was transfected into HEK293T cells with pEGFP-C1-Ccdc38, 48 hours after transfection, cells were collected for immunoprecipitation with anti-MYC, and detected by anti-GFP or anti-MYC antibodies, respectively. C CCDC146 co-localized with CCDC38 and γ-TUBULIN in Hela cells. pCSII-MYC-Ccdc146 and pEGFP-C1-Ccdc38 were co-transfected into Hela cells. 24 h after transfection, cells were fixed and stained with anti-MYC and anti-γ-TUBULIN or anti-GFP antibodies, and the nucleus was stained with DAPI. Scale bar: 5 μm. D The expression of Ccdc146 in different tissues of adult mice. GAPDH was used as a loading control. E The expression of Ccdc146 on different days. TUBULIN was used as a loading control. F The schematic illustration of the knockout strategy for generating Ccdc146−/− mice lacking exon 3–7. Primer-F1, Primer-R1, and Primer-R2 were used as the genotyping primers. G Genotyping of Ccdc146−/− mice. H Western blotting of CCDC146 indicated the depletion efficiency in Ccdc146−/− male mice. I Ccdc146−/− male mice were infertile. The Y axis represents the average number of offspring per litter. Data are presented as means ± SEM. two-tailed Student’s t test; ***P < 0·001. J The number of pups per litter of the different crossing possibilities. Data are presented as means ± SEM. two-tailed Student’s t test; ns: no significance; ***P < 0.001. K The sex ratio of the pups of the different crossing possibilities. Data are presented as means ± SEM. two-tailed Student’s t test; ns: no significance; ***P < 0.001. L The size of the Ccdc146+/+ and Ccdc146−/− mice testes were unaffected. M Testis from Ccdc146+/+ and Ccdc146−/− male mice had no obvious difference in weight (n = 5). Data are presented as means ± SEM. two-tailed Student’s t-test; ns: no significance. N The body weight of Ccdc146+/+ and Ccdc146−/− male mice had no obvious difference (n = 5). Data are presented as means ± SEM. two-tailed Student’s t-test; ns: no significance. O The testis/body weight ratios in Ccdc146+/+ and Ccdc146−/− male mice were consistent (n = 5). Data are presented as means ± SEM. two-tailed Student’s t-test; ns no significance