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. 2020 Jun 22;12(6):1862. doi: 10.3390/nu12061862

Figure 5.

Figure 5

Overexpression of claudin 3 rescues sweetener-induced barrier leakage across the intestinal epithelium. Caco-2 cells were transiently transfected with cDNA encoding wild-type claudin 3 (CLDN3-DYK) or the vector control (DYK), or untransfected (UT) for 48 h. (a) The total protein levels of claudin 3 in cells were measured by the Western blot analysis of cell lysates using 50 µg protein. (b) The cell surface expression of claudin 3 was determined by whole-cell indirect ELISA using chemiluminescence in transfected Caco-2 cells. (c) The viability of transfected Caco-2 cells was assessed by a CCK8 assay. Absorbance values were normalized to untransfected control and expressed as percentage viability. (d) The permeability of the transfected Caco-2 cell monolayer was measured by an FITC-dextran assay following exposure to sucralose and aspartame (0.1 mM) for 24 h. % permeability was calculated normalized to vehicle treatment. n = 6. Data are expressed as mean ± S.E.M. * p < 0.05 versus untransfected cells or vehicle (0 µM).