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. 2022 Aug 22;149(16):dev200499. doi: 10.1242/dev.200499

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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. 5. — Acrosome analysis using PNA-FITC fluorescence staining. (A-I) Immunofluorescence staining for acrosome biogenesis on testes sections and mature sperm of WT, Pfn4^+/− and Pfn4^−/− mice (n=3 biological replicates/genotype). In the Golgi phase, the proacrosomal granule (green) was labeled by PNA-FITC in WT (A), Pfn4^+/− (B) and Pfn4^−/− (C) round spermatozoa. In the cap phase, acrosomal caps (green) were stained in WT (D), Pfn4^+/− (E) and Pfn4^−/− (F) round spermatozoa (white arrows show abnormal cap structures). In the acrosomal phase, PNA-FITC-labeled the acrosomal area in WT (G), Pfn4^+/− (H) and Pfn4^−/− (I) elongated spermatids. Dashed boxes indicate the areas enlarged to the right. (J) Immunofluorescence staining using PNA-FITC (green) on epididymal sperm cells of WT, Pfn4^+/− and Pfn4^−/− mice (n=3/genotype) to show the variety of Pfn4^−/− sperm such as malformed acrosome and abnormal head morphology. Scale bars: 20 μm.