Figure 3.

Immunofluorescence assays of testicular germ cells from Ssp411^N-HA/N-HA mouse testes. (a) Western blotting analysis of various tissues from Ssp411^N-HA/N-HA mice using an anti-HA antibody. β-tubulin was used as the internal control. (b) Western blotting analysis of cauda spermatozoa isolated from the epididymis of Ssp411^N-HA/N-HA mice. α-tubulin was used as the internal control. (c) Statistical analysis of the grayscale values of Ssp411 relative to the internal control. Three replicated were performed. ^****P < 0.0001. (d) IF staining of testicular germ cells from Ssp411^HA/HA mice using an anti-HA antibody (HA-Ssp411, green), PNA (acrosome, red), and Hoechst 33342 (nucleus, blue). (e) IF staining of epididymal spermatozoa using anti-HA antibody (HA-Ssp411, green), Normal IgG was used as a negative control, and Hoechst 33342 (nucleus, blue) and TD (bright field) were used. Ssp411: sperm-specific protein 411; HA: hemagglutinin; PNA: peanut agglutinin; TD: transmitted light detection; IF: immunofluorescence.