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. Author manuscript; available in PMC: 2021 Mar 19.
Published in final edited form as: Cell. 2020 Mar 10;180(6):1160–1177.e20. doi: 10.1016/j.cell.2020.02.017

Figure 3. DDRGK1 Specifically Regulates ER-phagy.

Figure 3.

(A) ER-phagy CRISPRi screen identifies genes that are associated with the ER. Previously reported ER-phagy regulators are highlighted in red. DDRGK1 is highlighted in blue.

(B) DDRGK1 depletion results in inhibition of ER-phagy. HCT116 CRISPRi EATR cells were transduced with sgRNAs targeting ULK1 or DDRGK1 and starved for 16 h before FACS measurement for ER-phagy. Data are presented as mean ± SD of three biological replicates.

(C) DDRGK1 specifically inhibits ER-phagy but not general autophagy. HCT116 CRISPRi CCER cells were transduced with sgRNAs targeting DDRGK1 and starved for 16 h. Cells were lysed for western blotting of the indicated proteins.

(D) HCT116 CRISPRi cells stably expressing mCherry-eGFP-LC3B constructs were transduced with sgRNAs targeting either ULK1, ATG10, or DDRGK1. Cells were starved for 16 h before FACS measurement for general autophagy.

(E) DDRGK1 localizes to the ER. HeLa cells were stably transduced with DDRGK1-mCherry construct and immunostained for calnexin (CANX) as an ER marker. Insets represent a 3-fold enlargement of boxed areas. Scale bar represents 10 μm.

See also Figure S4.