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Fig. 9. Bgs4p localizes to the mating projections in an ordered mating type-dependent sequence, appearing first in P (h⁺) cells and later in M (h^–) cells. Early-logarithmic phase cells (GFP-bgs4⁺ bgs4 h^– and h⁺, and RFP-bgs4⁺ bgs4 h^– and h⁺) grown on EMM at 28°C were collected, washed and mixed in equal cell amounts (h⁺ GFP-bgs4⁺ x h^– RFP-bgs4⁺ and h⁺ RFP-bgs4⁺ x h^– GFP-bgs4⁺). Cell mixtures were transferred onto SPA plates and incubated at 28°C. Samples were collected at 30-minute intervals from 0 to 5 hours, ethanol-fixed, and examined for GFP and RFP fluorescence. Merged images of GFP (green) and RFP (red) fluorescence are shown. Arrowheads show the first time point at which h⁺ cells develop mating projections and localize Bgs4p (GFP or RFP). Arrows show the first time point at which mating projections and localized Bgs4p (GFP or RFP) are detected in h^– cells.

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