Journal
AUTOPHAGY
Volume 6, Issue 4, Pages 449-454Publisher
TAYLOR & FRANCIS INC
DOI: 10.4161/auto.6.4.11612
Keywords
autophagy; LC3; DOTAP; cationic lipids; mTOR; transfection
Categories
Funding
- Chinese Ministry of Sciences 973 Program [2006CB933300, 2009CB930300]
- Natural Science Foundation of China [30830036, 30721002]
- Chinese Academy of Sciences [KSCX2-YW-R-139, KSCX2-YW-R-21]
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Autophagy is a cellular stress response that results in the activation of a lysosomal degradation pathway. In this report, we showed that cationic lipids, a commonly used class of transfection reagents, induced genuine autophagy in mammalian cells. Extensive LC3 dot formation was observed by treatment with cationic lipids (with or without DNA), but not neutral lipids, in a HeLa cell line stably expressing GFP-LC3 (HeLa-LC3). Further proofs for autophagy were obtained by the colocalization of the LC3 dots with lysosome-specific staining patterns, observation of LC3-I to LC3-II form conversion and appearance of autophagic vacuoles under TEM. The autophagic flux assay with bafilomycin A1 and degradation of p62/SQSTM1 suggested that the autophagy induced by cationic lipids was primarily due to increased formation of autophagosomes and not decreased turnover. Moreover, cationic lipids induced autophagy in an mTOR-independent manner.
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