Figure 8. The turnover of both Hxt3 and Hxt7 is dependent on Rim15.

BY4742 (WT) and rim15Δ expressing MET25pro-HXT3-GFP were cultured in glucose media minus methionine as described in the methods (time 0). Following 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. BY4742 and rim15Δ expressing CUP1pro-GFP-HXT7 were cultured in raffinose media plus CuSO₄ as described in the methods (time 0). After harvesting and washing, the cells were resuspended in raffinose media devoid of CuSO₄ and treated with rapamycin. Samples were collected at the indicated times and analyzed by (C) fluorescence microscopy, and (B) Western analysis with anti-GFP antibodies. Identical blots were also probed with anti-ADH antibody as a loading control.