Fig. 3. Tea1p, Tip1p and Tea2p partially delocalise in the presence of pheromone. (A) tip1YFP-tea1GFP (B) and tip1YFP-tea2GFP strains were used to determine colocalisation in vivo. Cells were grown in minimal medium without or with pheromone for 5 (A) or 7 (B) hours. Pictures were taken with a confocal microscope using YFP and CFP filters, which allow excitation and detection of each fluorophore separately. The more narrow ends, marked by an asterisk, are known to be growing from previous calcofluor and actin stainings. The arrows in A indicate the Tea1GFP dots co-localising with Tip1YFP in the absence of pheromone but not in the presence of pheromone. In B the arrows indicate the Tea2GFP dots co-localising with Tip1YFP both in the presence and absence of pheromone. Scale bar, 3µm. (C) Images of a tea2GFP strain in the YFP and CFP channels, showing there is no bleed through in the YFP channel. Bar, 5 µm. (D) To confirm the localisation in a wild-type background, we mated an h90 Tea2GFP overnight on glutamate plates and photographed mating cells. Bar, 3 µm. (E) Images of Tea1GFP, Tip1YFP and Tea2GFP in the absence or after 6 hours in the presence of pheromone. Bar, 3 µm.