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. 2012 Jan 15;125(2):473–485. doi: 10.1242/jcs.093344

Fig. 7.

Fig. 7.

Decrease in [Na+]i is induced by genistein in non-capacitated sperm and blocked in the absence of Cl− in capacitated sperm. Sperm from the cauda epididymus placed in non-capacitating medium were loaded with CoroNa Red for 30 minutes, washed and then incubated for an additional hour in the same medium in the absence or presence of increasing concentrations of genistein. Another aliquot was incubated for 60 minutes in complete medium. (A–C) Two-dimensional dot plots of PI versus CoroNa Red fluorescence of sperm incubated in non-capacitating (NON; A), non-capacitating and 20 μM genistein (Gen; B) or in capacitating (CAP; C) media. (D) Normalized CoroNa Red fluorescence of live sperm that were incubated with increasing concentrations of genistein. Values are means ± s.e.m. of four independent experiments. (E) Western blots, using anti-PKAS-P and anti-Y-P, of sperm extracts incubated in medium that supported (CAP) or did not support (NC) capacitation, in the presence of 20 μM genistein. (F,G) Two-dimensional dot plots of PI versus CoroNa Red fluorescence of sperm in medium that supported capacitation, in the presence (CAP; F) or in the absence of Cl− (CAP, Cl− free; G). (H) The percentage of live sperm showing CoroNa Red fluorescence in CAP and CAP Cl−-free media. (I) Normalized CoroNa Red fluorescence of live sperm that were incubated in medium that supported capacitation, in the presence (CAP) or in the absence of Cl− (Cap Cl− free). Values are means ± s.e.m. of eight independent experiments (*P≤0.05; **P≤0.01; ***P≤0.001).