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. 2019 Mar 15;8(3):248. doi: 10.3390/cells8030248

Figure 1.

Figure 1

Nic could initiate non-canonical LC3 lipidation (NCLL), which is independent of the ULK1 complex and Beclin 1 complex, but dependent on the ubiquitin-like conjugation systems. (A) Wild-type MEFs (WT), FIP200KO-MEFs (FIP), and ULK1KO-MEFs (ULK) were treated with Nic (10 μM) for 6 h, and then, LC3-II formation was analyzed by immunoblotting. (B) Wild-type U251 cells (U251) and Beclin 1KD-U251 cells (Beclin 1) were treated with Nic (10 μM) for 6 h or EBSS for 2 h, then LC3-II formation was analyzed by immunoblotting. (C) WT-MEFs, FIP200KO-MEFs, ULK1KO-MEFs, and ATG5KO-MEFs stably expressing GFP-LC3 were treated with Nic (2.5 μM) for 6 h, and then, the GFP-LC3 puncta formation was analyzed. (D) Quantification of GFP-LC3 puncta in (C). (E) WT-MEFs and ATG5KO-MEFs were treated with Nic (10 μM) for 6 h, and then, the LC3-II formation was evaluated. (F) WT-HeLa cells (HeLa) and ATG4BKO-HeLa cells (ATG4B) were treated with Nic (10 μM) for 6 h and then analyzed by immunoblotting. CM, complete medium. ** p < 0.005. ns, not significant. Bar = 25 μM.