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. 2021 Oct 13;10(10):2730. doi: 10.3390/cells10102730

Figure 1.

Figure 1

PA-induced LAMP2 reduction, ER stress, and impaired autophagy flux is mTOR-dependent in hepatocytes. Hepa-1c1c7 cells, with or without transfection of tandem fluorescent mRFP-GFP-tagged LC3 plasmid (ptfLC3) were treated with 100 µM palmitic acid (PA) for 24 h in the absence or presence of a potent mTORC1 inhibitor rapamycin (50 nM). (A) Cellular LDH levels. (B) Immunoblot and quantification analysis of cellular protein levels of p-S6, S6, ATF4, CHOP, LAMP2, LAMP1, and LC3II. (C) Autophagy flux was visualized by immunofluorescence (IF) microscopy. (D) Quantification of LC3 dots in the cell. Red bars represent RFP-positive dots and yellow bars represent colocalization of RFP and GFP dots when merged. Scale bars, 20 μm. A one-way ANOVA test was performed to calculate the significance of the data (* p < 0.05). BSA, bovine serum albumin. PA, palmitic acid.