Figure 7.

Rapamycin treatment destabilizes Aly1 and Aly2, irrespective of Rsp5 binding: (A–D) Whole-cell extracts from WT cells treated with nothing (no treatment; NT), rapamycin (rapa 200 ng/mL), or cycloheximide (CHX 100 μg/mL) for the times indicated, and expressing (A) Aly1-GFP, (B) Aly1^PPXYless-GFP, (C) Aly2-GFP, or (D) Aly2^PPXYless-GFP, or from npr1∆ cells expressing (E) Aly1-GFP or (F) Aly2-GFP, were resolved by SDS-PAGE. Anti-GFP antibody was used to detect tagged α-arrestins and REVERT total protein stain was used as a loading control. Blots shown are one representative from 4 replicate experiments. Molecular weights are shown on the left side in kDa. For blots in panels (A–D), the pixel intensities for the GFP-detected band and the lane in the loading control were measured using Image J. A correction factor based on the loading control was applied to each pixel intensity measurement. The t = 0 point was set to 100%, and all abundances are presented relative to that point. The error bars represent the SEM.