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BY4742 (WT) and PGK1pro-VID28 expressing MET25pro-HXT3-GFP were cultured in glucose media minus methionine as described in the methods (time 0). Following washing and a switch to ethanol media plus methionine, samples were collected at the indicated times and analyzed by (A) fluorescence microscopy, and (B) Western analysis with anti-GFP antibodies. Identical blots were also probed with anti-ADH antibody as a loading control.
