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. 2012 May 15;23(10):1860–1873. doi: 10.1091/mbc.E11-09-0746

FIGURE 2:

FIGURE 2:

mAtg9 colocalizes with both early and late autophagosome autophagy markers. (A) Confocal microscopy of mAtg9 (red) in HEK293/GFP-LC3 and HEK293/GFP-DFCP1 cells. GFP-DFCP1, ULK1, WIPI2, Atg16, or GFP-LC3 in green as indicated. ULK1, WIPI2, Atg16, and mAtg9 were detected using antibodies as in Figure 1. Arrows in merge show mAtg9 colocalization with markers. Bars, 10 μm. (B) Confocal microscopy of HEK293/GFP-LC3 cells, GFP-LC3 (green), ULK1 (blue), mAtg9 (red); HEK293/GFP-DFCP1 cells with mAtg9 (red), LC3 (blue); HEK293/GFP-LC3 cells, GFP-LC3 (green), Atg9 (red), and WIPI2 and Atg16 (blue). A low and partial colocalization of ULK1 and Atg9 is shown and is in contrast to the colocalization of ULK1 with LC3. LC3 emerges from the GFP-DFCP1 structure, and mAtg9 coincides partially with both structures. Bottom, WIPI2, Atg16, and GFP-LC3 signals overlap, whereas mAtg9 is found in close proximity with WIPI2, Atg16, and GFP-LC3. Quantification of the percentage of Atg9 found with ULK1, DFCP1, WIPI2, Atg16, and LC3 is shown in Table 1.