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. 2022 Jan 14;149(2):dev199988. doi: 10.1242/dev.199988

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Fig. 2. — C2CD6 is essential for in vivo fertilization. (A) Targeted inactivation of the C2cd6 gene. The 1.6-kb deleted region includes exon 1 (containing the initiating start codon) and 600-bp upstream of exon 1 (presumably the promoter region). The neomycin selection marker PGKNeo is flanked by loxP sites. HyTK provides negative selection by ganciclovir in ESCs. (B) Western blot analysis of C2CD6 in adult testes. ACTB serves as a loading control. (C) Testis weight normalized to body weight of males at 3-6 months of age. (D) Sperm count of males at 3-6 months of age. NS, not statistically significant (unpaired Student's t-test). n=3. (E) In vivo fertilization assay. Embryos/unfertilized eggs were flushed from wild-type females mated with either C2cd6^+/− or C2cd6^−/− males.