FIGURE 7:

Pan1 mutants do not show significant defects in TORC2 signaling. (A) Wild-type and pan1-AID cells were grown at 30°C to log phase in YPD medium. Auxin (1 mM) was added, cells were collected at the indicated intervals, and Ypk-pT662 and Ypk1 protein levels were analyzed by Western blot. Quantification of Ypk-T662 phosphorylation and total Ypk1 protein are shown. (B) Wild-type and pan1-AID cells were grown in YPD medium to early log phase. Auxin (1 mM) was added, and after 30 min cells were washed into YPG/E medium at 30°C containing 1 mM auxin. Cells were collected at the indicated intervals, and Ypk-pT662 and Ypk1 protein levels were assayed by Western blot. (C) Cells of the indicated genotypes were grown in YPD medium to early log phase at 30°C, and Ypk-pT662 and Ypk1 protein were assayed by Western blot. An asterisk indicates a background band in the Ypk1 blot that overlaps with the Ypk1 signal. Error bars represent the SD of the mean of three biological replicates. *** indicates a p value smaller than 0.005, and ** indicates a p value between 0.01 and 0.005 when the indicated strain is compared with the wild type. * indicates a p value between 0.05 and 0.01.