Figure S2.
MDCs are not bound by autophagosomal membranes. (A–C) 2D cross sections and a 3D model derived from five serial 90-nm cell sections of the same yeast strain analyzed in Fig. 1. Sections were immunolabeled with antibodies targeting GFP and secondary antibodies conjugated to 10-nm gold particles. Scale bar = 200 nm. See associated Video 7. (A) Tomograph of a large, multilamellar structure strongly labeled with antibodies targeting Tom70-yEGFP. Yellow arrow: mitochondria, white arrow: multilamellar structure; white arrowheads point to gold particles. (B) Model overlay of the tomograph shown in A. (C) 3D model of a large, multilamellar structure bound by two closely apposed paired membranes. The limiting membrane of the outer doublet membrane is labeled green, while the internal doublet membrane is labeled cyan. Mitochondria: purple, ER: yellow; 10-nm gold particles are overlayed with green spheres. (D) Representative super-resolution confocal fluorescence microscopy images of yeast expressing Tom70-mCherry and sfGFP-Atg8 after a 2-h treatment with 200 nM Rap. White arrow marks MDC, orange arrow marks sfGFP-Atg8–positive structures. The images shown are from the same cell but show different z-slices, which are labeled. Scale bar = 1 µm. (E) Quantification of the frequency sfGFP-Atg8 foci were colocalized or closely associated with Tom70-mCherry–marked MDCs. Error bars show mean ± SE of three replicates, n ≥ 100 MDCs per replicate. (F–I) 2D cross sections and 3D models from different views of the same tomogram shown in Fig. 3, A–C. Scale bars = 200 nm. (F and G) Two different 2D cross sections from a larger view of the same tomogram are shown in Fig. 3, A–C. White arrow: MDC, yellow arrow: mitochondria, orange arrow: autophagosome. (H) Model overlay of the tomograph shown in F. (I) 3D model of a vacuole-associated autophagosome labeled in orange, MDC labeled in green, mitochondria labeled in purple, and the vacuole labeled in red.

MDCs are not bound by autophagosomal membranes. (A–C) 2D cross sections and a 3D model derived from five serial 90-nm cell sections of the same yeast strain analyzed in Fig. 1. Sections were immunolabeled with antibodies targeting GFP and secondary antibodies conjugated to 10-nm gold particles. Scale bar = 200 nm. See associated Video 7. (A) Tomograph of a large, multilamellar structure strongly labeled with antibodies targeting Tom70-yEGFP. Yellow arrow: mitochondria, white arrow: multilamellar structure; white arrowheads point to gold particles. (B) Model overlay of the tomograph shown in A. (C) 3D model of a large, multilamellar structure bound by two closely apposed paired membranes. The limiting membrane of the outer doublet membrane is labeled green, while the internal doublet membrane is labeled cyan. Mitochondria: purple, ER: yellow; 10-nm gold particles are overlayed with green spheres. (D) Representative super-resolution confocal fluorescence microscopy images of yeast expressing Tom70-mCherry and sfGFP-Atg8 after a 2-h treatment with 200 nM Rap. White arrow marks MDC, orange arrow marks sfGFP-Atg8–positive structures. The images shown are from the same cell but show different z-slices, which are labeled. Scale bar = 1 µm. (E) Quantification of the frequency sfGFP-Atg8 foci were colocalized or closely associated with Tom70-mCherry–marked MDCs. Error bars show mean ± SE of three replicates, n ≥ 100 MDCs per replicate. (F–I) 2D cross sections and 3D models from different views of the same tomogram shown in Fig. 3, A–C. Scale bars = 200 nm. (F and G) Two different 2D cross sections from a larger view of the same tomogram are shown in Fig. 3, A–C. White arrow: MDC, yellow arrow: mitochondria, orange arrow: autophagosome. (H) Model overlay of the tomograph shown in F. (I) 3D model of a vacuole-associated autophagosome labeled in orange, MDC labeled in green, mitochondria labeled in purple, and the vacuole labeled in red.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal