Fig. 4 | Journal of Cancer Research and Clinical Oncology

Fig. 4

From: Resveratrol induces PD-L1 expression through snail-driven activation of Wnt pathway in lung cancer cells

Fig. 4

Resveratrol-activated SirT1 deacetylates Snail and increases Snail stability. a Western blot demonstrates increased Snail expression following 3 h of resveratrol (0, 1, 5, 10, 20 and 30 μM) stimulation in different histological non-small-cell lung cancer cell lines. b RNA was extracted and subjected to qRT-PCR in A549 and H1299 cells with indicated resveratrol treatment. Values represent the relative reduction of Snail mRNA levels normalized to GAPDH. No significant differences were observed (difference from untreated cells by ANOVA with Dunnett’s correction for multiple comparisons was calculated). c A549 and H1299 cells were co-transfected with Snail promoter reporter and control Renilla luciferase reporter gene plasmid for 48 h and then treated with resveratrol for 3 h. Luciferase activity was determined and normalized using the dual luciferase reporter system. No significant differences were found. d The half-life of Snail was increased in cells treated with resveratrol compared with that in control cells, as demonstrated by CHX chase assays. Cells were exposed to CHX and collected at the indicated times. Cell lysates were immunoblotted with the indicated antibodies. e Western blot analysis of Snail in H1299 and A549 cells at day 3 after transfection with SirT1 cDNA or control vector and with resveratrol treatment. f Western blot analysis of Snail in the cells with resveratrol treatment at day 3 after transfection with wild-type SirT1 cDNA (SirT1/wt) or acetylase-dead SirT1 construct (SirT1/Mut) or empty vector. g Western blot analysis of Snail in the cells at day 3 after transfection with SirT1 siRNA or control siRNA. h Cells were transfected with SirT1 for 48 h, and then cell homogenates were immunoprecipitated with anti-Snail and blotted with anti-acetyl-lysine. i Co-immunoprecipitation assays were used to detect the direct interaction of SirT1 and Snail after transfection with SirT1 in the H1299 and A549 cells

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