Figure 6.

Effect of Emc10 deletion on ATPase expression and ionic equilibrium in spermatozoa. (A) Determination of ATP1A1, ATP1A4, and ATP1B3 protein levels in spermatozoa from wild-type (Wt) and Emc10-null (Ko) mice. α-tubulin was used as the protein loading control. The western blot shows a representative result of six independent experiments. (B) Quantitative analysis of ATP1A4 and ATP1B3 protein levels based on densitometry of western blot in A. (C) Intracellular Na⁺ concentration in spermatozoa from Wt and Ko mice. (D) Determination of intracellular pH in Wt and Ko spermatozoa incubated in HCO₃⁻-free medium or HCO₃⁻-containing medium for 0 or 1 h. (E) Determination of intracellular Ca²⁺ in Wt and Ko spermatozoa incubated in Ca²⁺-free or Ca²⁺-containing medium. (F) Detection of capacitation-associated protein tyrosine phosphorylation of Ko spermatozoa incubated in capacitating medium in the presence of 1 mM 8-Bromo-cAMP or 100 μM IBMX. A representative western blot of four independent experiments is shown. α-tubulin was used as the protein loading control. B, 8-Bromo-cAMP; I, IBMX. Data are presented as mean ± SEM, n = 4, **P < 0.01, ***P < 0.001, when compared with control.