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. 2019 Apr 2;100(6):1461–1472. doi: 10.1093/biolre/ioz052

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© The Author(s) 2019. Published by Oxford University Press on behalf of Society for the Study of Reproduction.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 5. — Indirect immunofluorescence analysis of PT proteins in mature rat spermatozoa, with accompanying perforatorial immunoblots. (A) Rows represent the localization pattern observed with indirect immunofluorescence in whole rat sperm with the following antibodies: WBP2NL, WBP2, GSTO2, and histone H3 (representative of core histones). DAPI is indicative of nuclear staining (blue), and antibody labeling is identified accordingly. Immunoreactivity with all antibodies tested was specific to the area of the perforatorium and postacrosomal sheath. The observed perforatorial labeling in falciform spermatozoa is a novelty not observed in spatulate sperm. (B) To confirm the perforatorial identity of these proteins, preparative immunoblots of isolated perforatorial fractions (see Figure 3A-3) were probed with each respective antibody and revealed immunoreactive bands characteristic of each protein. Bars, 10 μm.