Mechanism of CCCP-induced non-canonical autophagy / 中国生化药物杂志

ABSTRACT

Objective To identify the factors those regulate CCCP-induced non-canonical autophagy.Methods Different cells expressing GFP-LC3 were treated with or without CCCP (30μM) for 6 h.Fluorescent images were taken and cell lysates were analyzed by western blot assay.Real-time PCR was used to measure the mRNA levels of LC3B.FIP200-KO MEF cells were cultured and treated by 30 μM CCCP with or without water channel inhibitors, for 6 h.Cell lysates were analyzed by Western blot assay.Results CCCP could not induced autophagy in Atg5-KO MEF cells.CCCP could induce non-canonical autophagy in ULK1-KO MEF, FIP200-KO MEF, and Beclin1-KD U251.CCCP treatment in FIP200-KO MEF cells had no effect on the expression level of LC3B mRNA.We also found two distinct aquaporin water channel inhibitors could inhibit the generation of LC3 which was induced by CCCP.Conclusion CCCP induced non-canonical autophagy was Atg5-dependent, but Beclin1-, ULK1-and FIP200-independent.Osmotic imbalance could regulate CCCP-induce non-canonical autophagy.