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Fig. S1. Localization of Corona Red fluorescence and effect of mitochondrial uncoupling on Na+i. (A) CoroNa Red fluorescence distribution (left) and DIC and CoroNa Red merged images (right) of sperm incubated in medium that does not support capacitation. (B,C) Cauda epididymal sperm recovered in non capacitating medium was loaded with CoroNa Red for 30 minutes, washed and then incubated for an additional hour in the same medium. Five minutes before the beginning of the flow cytometry assay, 1 mM CCCP was added (C) or not (B) to sperm suspensions. Two-dimensional CoroNa Red vs PI fluorescence dot plots were used to analyze each condition. (D). Merged image of B and C, showing no significant difference between treatments. (E) Average values of CoroNa Red fluorescence of live sperm populations in NON and NON + 1 mM CCCP media. Results indicate means ± s.e.m., n=3.