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. 2021 Feb 6;22(4):1646. doi: 10.3390/ijms22041646

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Mitochondrial superoxide levels expressed as percentages of viable spermatozoa Mito-E⁺/YO-PRO-1⁻ (a,b), and peroxide levels expressed as percentages of viable spermatozoa DCF⁺/PI⁻, (c,d) in extended and frozen–thawed (FT) samples at 30 min and 240 min post-thaw. In Experiment 1 (n = 8), TEA, PAX, and 2-GBI blockers were added to LEY medium at 15 °C (a,c), whereas in Experiment 2 (n = 9), they were added to LEYGO medium at 5 °C (b,d). Different superscripts indicate significant differences (P < 0.05) between samples within the same time point. Results are given as mean ± SEM.