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. 2016 Jul 1;6:29045. doi: 10.1038/srep29045

Figure 5. Vacuole fusion upon mating.

Figure 5

(A) Plasmids expressing the PHO8 gene (ALP) fused to mCherry or GFP were introduced into isogenic a- and α-strains of BY4741 cells or the indicated nyv1Δ or vph1Δ mutants. a- and α-cells were grown over night in HC-Leu medium. In the morning, the cultures were diluted to OD600nm = 0.2, the respective a and α cells were mixed and incubated for 4–6 h at 30 °C. Upon emergence of the first diploid daughters (labeled by an asterisk), the cells were analyzed by spinning disc fluorescence microscopy. The frequency of daughter cells showing complete co-localization of mCherry and GFP in their vacuoles was determined from 90 cells (n = 3). (B) Vacuoles were isolated from the wildtype and nyv1Δ cells used in A. Their proteins were analyzed by SDS-PAGE and Western blotting against Vam3 and Nyv1.