Figure 6
HGF co-treatment prevents dedifferentiation of polarized Caco-2 cells after prolonged exposure to the VX-809/VX-770 drug combination. (a) Caco-2 colorectal cells were polarized to a TEER above 600 Ω, stained with phalloidin-TRITC, DAPI, and anti-CFTR/Alexa 488, and analysed by confocal microscopy. (b) Variation in TEER of polarized Caco-2 colorectal cells treated for 15 days with DMSO, HGF (50 ng/ml), VX-809 (3 μM), or VX-770 (0.5 μM), alone or in the indicated combinations. Conditions leading to significant TEER variations are indicated. (c) WB analysis of whole cell lysates from polarized Caco-2 cells treated as in (b). Shown are representative images of immunoblots for ZO-1, E-cadherin, CK8, and CK18 together with bar plots of band intensity quantification, normalized to DMSO (means ± SEM) from three independent assays. Histone H2B was used as a loading normalizer in band intensity quantification. (d–f) Immunofluorescence staining of polarized Caco-2 cells, treated as in (b), with phalloidin-TRITC, DAPI, and either anti-ZO-1/Alexa 488 (d), anti-E-cadherin/Alexa 488 (e), or anti-CK18/Alexa 488 (f) and analysed by confocal microscopy. Shown are overlay images representative of the indicated treatment conditions. White horizontal bars represent 10 μm. *p < 0.05; #p < 0.01.
