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Lactate produced by glycolysis increases the lactylation level of PKM2 protein, which inhibits its tetramer-to-dimer transition, reducing nuclear distribution and enhancing its pyruvate kinase activity in the cytoplasm (48)

After incubation with hypo-osmotic swelling solution [HOS, containing 6.75 g/L fructose and 3.67 g/L sodium citrate] at 37C for 1 h, plasma membrane integrity of each group (repeated in triplicated)was analyzed by evaluating the modifications of sperm tail to score swollen sperm according to previous publications (61)