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. 2020 May 4;21(9):3255. doi: 10.3390/ijms21093255

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© 2020 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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Percentages of viable sperm with high intracellular calcium levels evaluated through Fluo3 staining (Fluo3⁺) in relation to viable sperm (PI^-), and fluorescence intensity of Fluo3⁺ in viable sperm throughout incubation in capacitating medium (control samples) or capacitation medium plus 1 mM, 5 mM or 10 mM 2-GBI blocker, added at either 0 min in experiment 1 (A,B) or 240 min in experiment 2 (C,D; acute). Different superscript letters (a–c) indicate significant differences (p < 0.05) between samples within the same time point, and different superscript numbers (1–5) indicate significant differences (p < 0.05) between time points within a given treatment. Results are expressed as mean ± SEM (n = 10). The arrow indicates the time at which 10 µg/mL of progesterone was added to all samples (i.e., 240 min of incubation).