pex3 atg1 cells harbor Pex14-containing structures. (A) EM analysis of KMnO₄-fixed pex3 atg1 and WT cells grown for 16 h on MM-M/G. The inset shows a cluster of vesicles (enlarged from the boxed region). (B) iEM analysis of pex3 atg1 cells using α-Pex14 antibodies. (C and D) FM images of pex3 atg1 cells producing Pex14-mCherry and the ER marker BiP_N30-eGFP-HDEL (C), or Pex14-mGFP complemented with Mitotracker orange staining (D). (E) Electron tomography analysis of a serial-sectioned pex3 atg1 cell (a) containing a perinuclear membrane cluster (b, arrows). (d–f) 10-nm-thin digital slices through the tomogram reconstruction (viewing direction indicated in c) revealed vesicles (black arrows) and reticular structures (red arrows). The surface-rendered reconstruction in g shows the reticulovesicular structures in 3D viewed at right angles from d. CW, cell wall; M, mitochondrion; N, nucleus; P, peroxisome; V, vacuole. Bars: (A) 500 nm; (A, inset; and B) 100 nm; (C and D) 2.5 µm; (E) 250 nm.