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. 2019 Dec 20;7:319. doi: 10.3389/fcell.2019.00319

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Copyright © 2019 Nixon, Bernstein, Cafe, Delehedde, Sergeant, Anderson, Trigg, Eamens, Lord, Dun, De Iuliis and Bromfield.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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4HNE adduction of proAKAP4 and AKAP4 attenuates capacitation-associated signaling in human spermatozoa. Human spermatozoa treated with 4HNE (100 μM for 1 h at 37°C) were induced to capacitate (Capacitated) or held in a non-capacitated state (Non-cap) using standard protocols (Mitchell et al., 2007). Spermatozoa were subsequently prepared for either (A,B) immunocytochemistry or (C,D) immunoblotting with anti-phospho-PKA substrate or anti-phosphotyrosine antibodies. A densitometric trace of representative immunoblots is included to highlight changes in phospho-labeling in untreated (UT; black trace), GSH pre-treated (Pre-GSH; blue trace) and 4HNE treated (4HNE; red trace) spermatozoa. This experiment was replicated with samples from three separate donors and representative images are immunoblots are presented. Scale bars = 10 μm.