Atg37 binds and requires Atg30 for proper localization. (A) Atg37 coimmunoprecipitates with Atg30. Methanol-grown cells were adapted to glucose medium for 0.5 h to induce pexophagy. (B) Atg37ΔC (BD-37ΔC) interacts with Atg30 (AD-30) in Y2H. 3-AT, 10 mM 3-amino-1,2,4-triazole; AD, activation domain; BD, DNA binding domain. (C and D) Atg30 does not bridge the interaction between Pex3 and Atg37. Pex3 still coimmunoprecipitated with Atg37 in the absence of Atg30 under both peroxisome proliferation (C) and pexophagy (D) conditions. (E) Atg37 is not required for peroxisomal localization of Atg30. Cells were grown in methanol medium. Bar, 5 µm. (F) Atg30 is required for localization of Atg37 at the peroxisome cluster periphery. Arrows point to the peroxisome cluster with an accumulation of Atg37-GFP in the middle. Bar, 5 µm. (G) Quantitation of data in F presented as the mean + SD (error bars; n = 3; *, P < 0.001 vs. WT). (H) Atg37 accumulates in the middle of the peroxisome cluster in Δatg30 cells. Cells were grown in methanol medium. Z slices of peroxisome clusters are labeled 1–10. Bar, 2 µm.