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. 2016 Aug 11;7:12420. doi: 10.1038/ncomms12420

Figure 5. Combination of ERES-ATG9-VMP1 compartments can only partially predict the site of autophagosome nucleation.

Figure 5

(a) HEK293 cells stably expressing GFP-ATG13 were fed or starved, immunolabelled for ATG9 and ERGIC53 and imaged by wide-field microscopy. Bar corresponds to 10 μm. (b,c) Wide-field live-cell imaging of starved HEK293 cells expressing stably GFP-ATG13 and transiently VMP1-mCherry. Representative montages of ATG13 particles forming in association with VMP1 (b) or not (c) are shown. Arrowheads point at the ATG13 particles in the first two frames from their emergence, the same that were used for the analysis in f. (d) HEK293 cells transiently expressing VMP1-YFP were immunolabelled for ubiquitin and imaged by wide-field microscopy. Representative images are shown. Bar corresponds to 10 μm. (e) Wide-field live-cell imaging of starved HEK293 cells expressing stably GFP-ATG13 and mRFP-ATG9 and transiently CFP-SEC16 and VMP1-LSS-mKate2. Representative images of a cell expressing the four proteins are shown. Bar corresponds to 10 μm. (f) Values are ATG13 particles in b,c associating with VMP1 in the first two frames from their emergence. From analysis of 67 montages. (g,h) Values are ATG13 particles associating with any of ATG9, SEC16 and VMP1 (g) or with different combinations of them (h) in the first two frames from their emergence. Bar in b,c corresponds to 300 nm.