Fig. 6: Eukaryotic initiation factor 2α (eIF2α) is essential for pterostilbene (PT)-induced autophagy. | Cell Death & Disease

Fig. 6: Eukaryotic initiation factor 2α (eIF2α) is essential for pterostilbene (PT)-induced autophagy.

From: Inhibition of eIF2α dephosphorylation accelerates pterostilbene-induced cell death in human hepatocellular carcinoma cells in an ER stress and autophagy-dependent manner

Fig. 6

a Huh-7 cells treated with PT for 24 h were fixed; permeabilised; stained with anti-p-eIF2α antibody, anti-LC3 antibody and 4ʹ,6ʹ-diamidino-2-phenylindole (DAPI); and then visualised under a confocal microscope. b Cytoplasmic and nuclear proteins were isolated and the p-eIF2α, LC3-II and ATF4 protein expression levels were measured using Western blot analysis. Lamin B: nuclear marker; α-tubulin: cytoplasmic marker; and calreticulin: ER marker. c Huh-7 cells were treated with PT (100 μM) in the presence or absence of si-eIF2α for 24 h. The protein expression level was assessed using Western blot analysis with β-actin serving as an internal control. d Cell viability was measured using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. e Formation of acidic vesicular organelle (AVOs) was visualised through acridine orange (AO) staining and observed under a fluorescence microscope. f Quantification was performed using a flow cytometer. Data are presented as mean ± standard error for three experiments. **p < 0.01 compared with controls. #p < 0.01 compared with PT treatment alone

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