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Figure 1: Bile acids induce nuclear factor-kappa B in esophageal epithelial cells. (a) Effect of bile acids on nuclear factor-kappa B DNA-binding activity. OE33 cells were treated with bile acids deoxycholic acid, cholic acid, chenodeoxycholic acid, lithocholic acid and ursodeoxycholic acid at 300 μM for 2 h, nuclear extracts were prepared and analyzed by gel shift assay (as described under Experimental Procedures Section). (b) Effect of bile acids on IκB-α protein levels. OE33 cells were treated with 300 μM deoxycholic acid, cholic acid, chenodeoxycholic acid, lithocholic acid or ursodeoxycholic acid and Western blot analysis for IκB-α detection were performed. β-actin was used as a loading control. Experiments were performed 3 times with similar results and a representative gel is shown. High content analysis of nuclear factor-kappa B nuclear translocation by bile acids in OE33 cells (c) and SKGT-4 cells (d). SKGT-4 or OE33 cells were treated with different bile acids (deoxycholic acid, cholic acid, chenodeoxycholic acid, cholic acid or ursodeoxycholic acid) at 300 mM for 2 h. Mean from 3 wells of the difference in nuclear factor-kappa B fluorescence intensities (CircRingAvgIntenDiffCh2) is shown

Figure 1: Bile acids induce nuclear factor-kappa B in esophageal epithelial cells. (a) Effect of bile acids on nuclear factor-kappa B DNA-binding activity. OE33 cells were treated with bile acids deoxycholic acid, cholic acid, chenodeoxycholic acid, lithocholic acid and ursodeoxycholic acid at 300 μM for 2 h, nuclear extracts were prepared and analyzed by gel shift assay (as described under Experimental Procedures Section). (b) Effect of bile acids on IκB-α protein levels. OE33 cells were treated with 300 μM deoxycholic acid, cholic acid, chenodeoxycholic acid, lithocholic acid or ursodeoxycholic acid and Western blot analysis for IκB-α detection were performed. β-actin was used as a loading control. Experiments were performed 3 times with similar results and a representative gel is shown. High content analysis of nuclear factor-kappa B nuclear translocation by bile acids in OE33 cells (c) and SKGT-4 cells (d). SKGT-4 or OE33 cells were treated with different bile acids (deoxycholic acid, cholic acid, chenodeoxycholic acid, cholic acid or ursodeoxycholic acid) at 300 mM for 2 h. Mean from 3 wells of the difference in nuclear factor-kappa B fluorescence intensities (CircRingAvgIntenDiffCh2) is shown