Figure 1
From: Golgi-associated LC3 lipidation requires V-ATPase in noncanonical autophagy
AMDE-1 can induce NCA, which is independent of FIP200 and ULK1 but dependent on the conjugation system. (a) Wild- type MEFs (WT), FIP200KO-MEFs (FIP), ULK1KO-MEFs (ULK) and Atg5KO-MEFs (Atg5) were treated with AMDE-1 (10 μM) for 6 h, or EBSS for 2 h, and then analyzed by immunoblotting. (b) Immunoblotting analysis of WT-MEFs and FIP200KO-MEFs treated by AMDE-1 (10 μM) at different time points. (c) WT-MEFs, FIP200KO-MEFs, ULK1KO-MEFs and Atg5KO-MEFs expressing GFP-LC3 were treated with or without AMDE-1 (10 μM) for 6 h, or EBSS for 2 h and then assessed for LC3 puncta formation (green). (d) Qualification of GFP-LC3 puncta in (c). (e) FIP200KO-MEFs expressing GFP-LC3G120A were treated with or without AMDE-1 (10 μM) for 6 h and then assessed for LC3 puncta formation (green). ***P<0.001; NS, not significant. Bar=25 μm
