Fig. 5.

Atg1 and Atg13 interaction is unaltered in the absence of Ptc2 and Ptc3 phosphatases. (A) Atg1-Atg13-Atg17 binding in WT and ptc2Δ ptc3Δ strains before and after 4-h rapamycin treatment. Samples taken from cell lysates (Left) were blotted for anti-MYC to detect Atg13, or with anti-GFP or anti-Pgk1. Samples collected following Atg13-13×MYC immunoprecipitation (Right) were blotted for anti-FLAG to detect Atg1, anti-MYC to detect Atg13, and anti-Atg17. (B) Normalized Atg1-Atg13 binding was calculated as a ratio of FLAG-Atg1 band intensity to Atg13-MYC band intensity at 0 h for WT and ptc2Δ ptc3Δ and then normalized to WT. (C) Normalized Atg13-Atg17 binding was calculated as a ratio of Atg17 band intensity to Atg13-MYC band intensity at 0 h for WT and ptc2Δ ptc3Δ cells and then normalized to WT.