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Fig. 2 | Journal of Experimental & Clinical Cancer Research

Fig. 2

From: APE1 redox function is required for activation of Yes-associated protein 1 under reflux conditions in Barrett’s-associated esophageal adenocarcinomas

Fig. 2

ABS induces YAP1 expression, promotes its nuclear accumulation and facilitates its transcriptional activity. A Western blot analysis of APE1 and YAP1 levels in FLO-1 and OE33 cells. Cells were exposed to 200 µM ABS for 20 min followed by recovery in complete media for the indicated time points. β-actin was used as a loading control. B Representative immunofluorescent staining images of APE1 (red) and YAP1 (green) in FLO-1 cells exposed to ABS (200 µM, pH 4, 20 min) followed by 3 h recovery versus control untreated cells. DAPI (blue) was used for nuclear staining. Quantification of stain intensity by ImageJ is also reported. C Representative immunofluorescent staining images of APE1 (red) and YAP1 (green) in FLO-1 cells repeatedly exposed to ABS (200 µM, pH 5.5, 20 min per day for 14 days) versus control untreated cells. DAPI (blue) was used for nuclear staining. Quantification of stain intensity by ImageJ is also reported. D-E YAP1 transcriptional activity was measured using 8xCTIIC luciferase reporter assay in FLO-1 cells exposed or not to ABS treatment (200 µM, pH 4, 20 min) followed by 3 h recovery (D) or FLO-1 cells repeatedly exposed to ABS (200 µM, pH 5.5, 20 min per day for 14 days) versus control untreated cells (E). The luciferase reporter activity values were normalized to Renilla expression levels. F qRT-PCR analyses of YAP1 downstream target genes, CTGF, CYR61 and ANKRD1 in FLO-1 cells exposed to ABS (200 µM, pH 4, 20 min) followed by 3 h recovery versus control untreated cells. Values are represented as mean ± SEM of three independent experiments. * P < 0.05; ** P < 0.01; *** P < 0.001. G Representative immunohistochemistry staining images of APE1 and YAP1 in L2-IL-1β transgenic mouse tissues with high-grade dysplasia (HGD) and esophageal adenocarcinoma (EAC) lesions (Untreated: 8mice; HGD and EAC: 8mice each). Quantification of stain intensity by ImageJ is shown

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