Fig. 2 | Journal of Cancer Research and Clinical Oncology

Fig. 2

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

Fig. 2

β-catenin/TCF transcriptional complex is required for resveratrol-induced PD-L1 expression. a Western blot analysis of Axin2 and β-catenin in H1299 and A549 cells treated with indicated amount of resveratrol, and it showed that Wnt pathway is activated by resveratrol. b Western blot analysis of Axin2, β-catenin, and PD-L1 in indicated cells at day 3 after transfection with Wnt3A cDNA or empty vector and then treated with resveratrol (upper panel) and the quantification of each protein levels by densitometry were shown in lower panel. c Western blot analysis of Axin2, β-catenin and PD-L1 in A549 and H1299 cells treated with XAV-939 at a concentration of 10 μM for 1 h and then with different concentrations resveratrol for 3 h (upper panel) and quantification was in lower panel. d Scheme of PD-L1 luciferase reporter constructs containing three potential TCF-4 binding sites (1011) with different mutations (1011 M, 1011 M2 and 1011 M3). Mutations were made in sequence at three potential TCF-4 binding sites (1011 M1: CCAAAG to CCTTTG, 1011 M2: TAAAAG to CATTTT on the construct of 1011 M1 and 1011 M3: AAAAAA to CATTTT on the construct of 1011 M2). e The cells were transiently transfected with different PD-L1 promoter reporters for 48 h and treated with resveratrol for 3 h. Luciferase activity was determined and normalized using the dual luciferase reporter system (****p < 0.001 for the difference from the untreated reporter-transfected cells by ANOVA with Dunnett’s correction for multiple comparisons). f ChIP assays were performed using anti-β-catenin antibodies. The standard PCR product is run and scan (left panel). Quantitative RT-PCR results were expressed using histograms (right panel). The bars represent the mean ± SD of triplicates (**p < 0.01 for difference from empty control plasmid-transfected cells by ANOVA for multiple comparison)

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