Mitochondrial protein markers do not localize to the Atg9 clusters. (A) Ultrathin cryosections obtained from a wild-type (MMY067) strain were immunogold labeled only for Por1 because the anti-Por1 antibody does not work in double labeling. The broken line highlights an Atg9 cluster. (B–F) Cells expressing both Atg9-GFP and the mitochondrial marker Idh1-3xHA (MOY003) were processed for IEM, and cryo-sections were double immunogold labeled for GFP and HA. CW, cell wall; M, mitochondria; N, nucleus; PM, plasma membrane; V, vacuole. Bars, 200 nm.