Fig. 5: Pterostilbene (PT) induces activating transcription factor-4 (ATF4) binding to the LC3 promoter, which mediates cell death. | Cell Death & Disease

Fig. 5: Pterostilbene (PT) induces activating transcription factor-4 (ATF4) binding to the LC3 promoter, which mediates cell death.

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

Fig. 5

a Huh-7 cells treated with PT (100 μM) for 24 h were fixed; permeabilised; stained with anti-ATF4 antibody, anti-LC3 antibody and 4ʹ,6ʹ-diamidino-2-phenylindole (DAPI); and then visualised under a confocal microscope. b Nuclear proteins were isolated and assessed using Western blot analysis with Lamin serving as an internal control. c The binding sites of ATF4 transcription factor on the LC3 promoter are labelled. d Chromatin immunoprecipitation (ChIP) assay using anti-ATF4 antibodies was performed with DNA samples extracted from PT-treated Huh-7 cells. Input samples were used as positive controls. e Huh-7 cells were treated with PT (100 μM) in the presence or absence of siATF4 for 24 h. The protein expression levels were assessed using Western blot analysis. β-Actin served as an internal control. f Quantitative analysis of acridine orange (AO)-stained cells was performed using a flow cytometer. g Cell viability was measured using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. 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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