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This is due in part to the use of fluorescent resonance energy transfer (FRET), fluorophores were carefully selected such that, when the fluorophores are paired, activation of the donor fluorophore causes a highly efficient energy transfer to the acceptor fluorophore, producing a large emission from the acceptor fluorophore, this effect diminishes exponentially as the distance between the fluorophores increases

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Immunofluorescence staining of oocytes After a brief rinse with PBS/0.05% polyvinylpyrrolidone (to prevent adhesion), the oocytes were permeabilized with 0.5% Triton X-100/PHEM for 5 min, fixed in 3.7% PFA/PHEM (18.14 g PIPES, 6.5 g HEPES, 3.8 g EGTA, 0.99 g MgSO4, pH 7.0 w/KOH) for 20 min, and washed three times (10 min each) with PBS/polyvinylpyrrolidone