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. 2024 Jan 10;223(3):e202307147. doi: 10.1083/jcb.202307147

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© 2024 Hoque et al.

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Figure 3. — The annulus is displaced into the principal piece in Cby3^−/− sperm. (A) Cauda epididymal sperm were immunostained for SEPT4 and AKAP4. Asterisks indicate the MP/PP junction. (B) Quantification of sperm with the indicated annulus positions (n = 8 fields of view with ≥ 25 annuli per field). (C) SEM and TEM studies of cauda epididymal sperm. Enlarged images of the boxed regions are shown in the middle panels. Arrowheads denote the annulus. Asterisks indicate the MP/PP junction. MP, midpiece; PP, principal piece; CD, cytoplasmic droplet. (D) Testis lysates prepared from adult mice were processed for Western blotting using antibodies against SEPT2, 4, and 12. GAPDH was used as a loading control. (E) Quantification of the distance from the base of the flagellum to the distal end of the MP or the annulus (n = 8 fields of view with ≥60 sperm per field). Mitochondria and the annulus were detected by Biotracker and SEPT4 antibody, respectively. (F) Cauda epididymal sperm were immunostained for CD147 and SEPT12 (arrowheads). Nuclei were detected with DAPI. Scale bars for A, 10 and 5 µm (enlarged images); C, 1 µm and 200 nm (enlarged images); F, 10 μm. Means ± SD are shown. Student’s t test: ***P < 0.001, not significant (ns). Source data are available for this figure: SourceData F3.