Skip to main content
. 2011 Jul 22;146(2):303–317. doi: 10.1016/j.cell.2011.06.023

Figure 5.

Figure 5

N-ethylmaleimide Treatment Decreases Atg16L1 Precursor Maturation

(A) HeLa cells transiently transfected for 20 hr with Atg16L1-Flag and Atg5-GFP were treated as indicated with N-ethylmaleimide (NEM; 100 μM) for 10 min. Representative confocal pictures are shown. Scale bars, 5 μm.

(B) Size distribution of Atg16L1-Flag vesicles from (A). n > 40 cells for untreated and NEM-treated cells; three independent experiments.

(C) Size distribution of Atg5-GFP vesicles from (A). n > 20 cells for untreated and NEM-treated cells from three independent experiments.

(D) HeLa cells were treated as indicated with N-ethylmaleimide (NEM; 100 μM) for 10 min. Cells were immunolabeled for endogenous Atg16L1 and LC3 and subjected to confocal analysis. Scale bars, 5 μm.

(E) Colocalization efficiency between Atg16L1 and LC3 from (D). At least 50 cells were counted per experiment; data are the mean ± SD of the percentage of Atg16L1 vesicles containing LC3; three independent experiments.

(F) HeLa cells were treated as indicated with N-ethylmaleimide (NEM; 100 μM) for 10 min. Cells were immunolabeled for endogenous Atg16L1 and Atg12 and subjected to confocal analysis. Scale bars, 5 μm. At least 50 cells were counted per experiment and the data are mean ± SD of the number of Atg16L1 or Atg12 vesicles per cell; three independent experiments.

See also Figure S5.