Fig. 4.

Disorganized flagellum and manchette in Ccdc146^−/− spermatids. A The histology of the seminiferous tubules from Ccdc146^+/+ and Ccdc146^−/− male mice. B Immunofluorescence analysis of Ac-TUBULIN (red) and PNA lectin (green) to identify sperm flagellum biogenesis. C Abnormal manchette elongation in Ccdc146^−/− spermatids. Spermatids from different manchette-containing steps were stained with α/β-TUBULIN antibody (red) to visualize the manchette. Ccdc146^−/− spermatids displayed abnormal elongation of the manchette. D Statistical analysis of the length of the manchette. The color columns indicate the manchette length of the Ccdc146^−/− spermatids in different steps, while the black columns indicate the manchette length of the Ccdc146^+/+ spermatids. Data are presented as means ± SEM. two-tailed Student’s t test; ns no significance; ***P < 0.001. E TEM revealed that the manchette of elongating spermatids (steps 9–11) from Ccdc146.^−/− mice were ectopically placed. The red arrowhead indicates the abnormal manchette. Scale bars: 100 μm (A); 50 μm (A); 20 μm (B); 15 μm (A); 10 μm (B); 5 μm (C); 2·5 μm (E)