Figure 5
Impaired de novo DNA methylation in Snf5 CKO male germ cells. (A) Immunohistochemistry of the E17.5 testes using antibodies against DNMT3L, DNMT3A, MILI, and MIWI2. Three mice were analyzed for each genotype, and the mean ± standard deviation is shown. The nuclei were counterstained with DAPI. The percentages of germ cells expressing these proteins were significantly lower in the Snf5 CKO males (**P < 0.01, ***P < 0.001, Student’s t-test). Note that MIWI2 was absent from the nuclei and localized in the cytoplasm in some germ cells of the Snf5 CKO males (yellow arrows). C cytoplasmic localization; N nuclear localization. Bar: 10 µm. (B,C) The DNA methylation status of retrotransposons and imprinted genes in male germ cells. The DNA methylation levels of IAP, LINE-1 (type A, type Tf) (B), and H19-DMR and IG-DMR (C) were examined by BS-Seq analyses using the purified germ cells at E17.5–E19.5.
