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. 2022 Jun 17;149(12):dev200330. doi: 10.1242/dev.200330

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© 2022. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 6. — Sperm basic protein analysis in protamine-deficient sperm. (A) Representative images of CMA3 staining of Prm1^+/+, Prm1^+/− and Prm2^+/− epididymal sperm heads taken at the same exposure time. DAPI was used as a counterstain. (B) Mean percentage of CMA3-positive sperm in Prm1^+/−, Prm2^+/− and WT males (n=3). A minimum of 400 sperm per male were analyzed. (C) Representative AU-PAGE of basic protein extractions from WT, Prm1^+/−, Prm1^−/−, Prm2^+/− and Prm2^−/− epididymal sperm. Non-protamine basic proteins were detected at the top of the AU-PAGE. PRM1 and PRM2 ran at the bottom of the gel. Pre-PRM2 ran higher than did mature PRM (dashed vermillion box). (D) Percentage of mPRM2 of PRM in basic protein extractions from WT, Prm1^+/− and Prm2^+/− epididymal sperm. (E) Percentage of total PRM2 (including pre-PRM2) of PRM in basic protein extractions from WT, Prm1^+/− and Prm2^+/− epididymal sperm. (F) Percentage of pre-PRM2 of PRM2 in basic protein extractions from Prm1^+/−, Prm1^−/− and Prm2^+/− epididymal sperm. Data are mean±s.d. and were analyzed using a two-tailed, unpaired Student's t-test (*P<0.05; **P<0.005; ***P<0.001). Scale bars: 20 µm.