Fig. 2

Acidic bile salts induce IGFBP2 in esophageal adenocarcinoma cells. FLO1 and OE33 cells were treated with acidic bile salts (ABS, pH 4, 200 μM) for 20 min and then recovered in regular medium for designated time points. a and b, Western blotting analysis for IGFBP2 protein expression in FLO1 (a) and OE33 (b) cells. c and d, qRT-PCR for IGFBP2 mRNA expression in FLO1 (c) and OE33 (d) cells. e and f, immunofluorescence staining of IGFBP2 protein (red) in 3D organotypic cultures of FLO1 (e) and OE33 (f) cells. DAPI was used to stain nucleus (blue). ABS, acidic bile salts; UT, untreated with ABS; *** p < 0.001, ** p < 0.01, * p < 0.05