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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)

Further examination of these TM3 kink regions allowed us to identify a point mutation, Tc MscS C66L, that demonstrated higher tension sensitivity (Supplementary Fig

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