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. 2018 Jan 9;37(4):e97840. doi: 10.15252/embj.201797840

Figure 2. The WD domain of ATG16L1 is not required for canonical autophagy.

Figure 2

  • A
    Diagram of full‐length (FL) 229–242 deletion (ΔFBD) and 1–336 (ΔWD) ATG16L1 constructs used in this study.
  • B‐D
    Western blot analysis of ATG16L1 in (B) HCT116 ATG16L1 −/−, (C) MEF Atg16L1 −/− and (D) MCF10A ATG16L1 −/− cells stably re‐expressing ATG16L1 constructs. Arrows indicate specific ATG16L1 band.
  • E
    Western blotting for LC3 in complemented HCT116 cells ± PP242 (1 μM, 1 h).
  • F
    Quantification of fold differences of LC3‐II/LC3‐I ratios over controls from (E).
  • G
    Confocal images of GFP‐LC3 in complemented MEF cells ± starvation (1 h). Scale bar: 10 μm.
  • H
    Quantification of GFP‐LC3 puncta from 100 MEF cells per experiment cultured in full media (control) or EBSS (starve) for 1 h.
  • I
    Quantification of WIPI2b puncta in ATG16L1‐complemented HCT116 cells. Puncta from 100 cells were counted per experiment.
Data information: Data represent mean ± SEM from three separate experiments. (F) *P < 0.02. (H) ***P < 0.0001, **P < 0.001. (I) ***P < 0.0006, **P≪0.005 (Student's t‐test).