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. Author manuscript; available in PMC: 2013 Oct 1.
Published in final edited form as: Growth Factors. 2012 Sep 5;30(5):320–332. doi: 10.3109/08977194.2012.717076

Figure 3.

Figure 3

EGFR ligands reduce TER of polarized Caco-2 cells and AREG is detected in CM from Caco-2 and 18Co cells. (A) Polarized Caco-2 cells were treated with 10 ng/ml of EGF, TGFα, AREG, or HB-EGF on day seven after plating on Transwell® filters (arrow). All of the EGFR ligands tested significantly decreased Caco-2 TER (p < 0.001). Data shown are mean ± S.E. of three independent experiments. (B) EKI-785 (1μM), an irreversible EGFR kinase inhibitor, was added to polarized Caco-2 cells one (squares) or seven (triangles) days after plating on Transwell® filters. Addition of EKI-785 increased Caco-2 TER compared to CTL medium under both experimental conditions. Data shown are mean ± S.E. of three independent experiments. *†p < 0.001, compared to TER in CTL medium. (C) CM from Caco-2 cells, 18Co cells, and Caco-2/18Co co-cultured cells was collected, and AREG levels were determined by ELISA. When Caco-2 cells were grown in serum-free medium, AREG levels were seven-fold greater than that in CM from 18Co cells grown in serum-free medium. AREG levels detected in 18Co CM were increased by 13-fold from cells cultured in 5% serum-containing medium, compared to serum-free medium. Data shown are mean ± S.E. of three independent experiments. *p < 0.001, compared to that in serum-free 18Co CM.