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. 2017 Jan 27;8:14182. doi: 10.1038/ncomms14182

Figure 2. BCAS2 is required for germ cell development and male fertility.

Figure 2

(a) Real-time RT-PCR analysis of Bcas2 mRNA levels in control and Bcas2F/−;Vasa-Cre (Bcas2F/−;V-cre) testes of P8 mice with Hprt as the internal control (n=3). Error bars represent s.e.m. (b) Western blotting analysis of BCAS2 protein in control and Bcas2F/−;Vasa-Cre testes of P8 mice. MVH and α-tubulin was used as a germ cell marker and loading control, respectively. (c) Relative abundances of BCAS2 in the control and Bcas2F/−;Vasa-Cre testes of P8 mice were determined by Western blotting analyses from 4 independent experiments. Error bars represent s.e.m. (d) IF staining of BCAS2 in the control and Bcas2F/−;Vasa-Cre testes of P8 mice. White circles denote the BCAS2 null spermatogonia. PLZF was co-stained to indicate the location of spermatogonia. The DNA was stained with Hoechst 33,342. Scale bar, 20 μm. (e) Morphological analysis of adult testes showed that the Bcas2F/−;Vasa-Cre testes were smaller than the control. (f) Testes weight of adult control and Bcas2F/−;Vasa-Cre mice (***P<0.001, n=5). Error bars represent s.e.m. (g) Hematoxylin and eosin (H&E) staining of adult testes in control and Bcas2F/−;Vasa-Cre mice. Spermatocytes and spermatids were almost absent in the seminiferous tubules of the Bcas2F/−;Vasa-Cre mice. Scale bar, 100 μm. (h) IF staining of MVH (a germ cell marker) in adult testes in control and Bcas2F/−;Vasa-Cre mice. Compared with the control, only a few MVH positive cells in the basement membrane were observed in Bcas2F/−;Vasa-Cre testes. The DNA was stained with Hoechst 33342. Scale bar, 50 μm.