ATG gene requirements for GTA. (A) ATG deletion strains expressing GFP-Atg8 were treated with 0.04% MMS for 4 h, and GFP immunoblots were performed. Representative blots are shown in A, and all other blots are shown in SI Appendix, Fig. S3A. Results of all ATG mutants tested are summarized in SI Appendix, Table S1. (B) GTA does not induce canonical selective autophagy pathways. WT cells expressing either GFP-Atg8 or Om45-GFP were treated with MMS (0.04%) or rapamycin (200 ng/mL) for 4 and 12 h, respectively. GFP immunoblot was performed to monitor GTA and mitophagy. (C) Strains expressing various GFP fusions as indicated were treated with MMS to monitor different forms of selective autophagy. (D) GTA does not induce macroautophagy. The Pho8∆60 assay was performed on MMS-treated WT and atg7Δ cells expressing pho8Δ60. (E) Analysis of prApe1 processing after rapamycin or MMS. Strains of the indicated genotypes were treated with either rapamycin or MMS, and prApe1 processing was judged by immunoblot. Error bars reflect SD from three independent experiments.