Figure 4

HGF and TGFβ1 oppositely influenced the patterns of MET/EMT genes in 1833 and MDA-MB231 cells. a, b and c Representative images of Western blots performed with total protein extracts from 1833 and MDA-MB231 cells. The experiments were repeated three times with similar results. The samples were run on gel, and were processed under the same experimental conditions. Vinculin was used for normalization. For a and b, the numbers at the bottom indicate the fold variations relative to the starvation value, considered as 1; for c, the E-cadherin value for MDA-MB231 cells was compared to that of 1833 cells. For MMP2, the fold variations were calculated for the active form. d The cells, exposed to 1-h HGF on coverslips, were probed with anti-E-cadherin antibody (red); the nuclei were stained with DAPI, and their merge images are shown. All the images, taken at 400× magnification, are representative of experiments performed in triplicate. e Transcription-factor binding sites present in the entire E-cadherin promoter are shown. The 1833 cells, transiently transfected with the construct containing the entire E-cadherin promoter, were co-transfected with Twist-dominant negative (ΔTwist), WWOX e.v. or siRNAWWOX in the presence or the absence of HGF and TGFβ1. The histograms indicate the absolute values of Firefly/Renilla luciferase activity ratios. The data are the means ± S.E. of three independent experiments performed in triplicate. *P < 0.05, **P < 0.005 versus the basal luciferase activity value; ^ΔP < 0.05, ^ΔΔP < 0.005 versus the luciferase activity value in response to HGF