Figure 1

TM-Grb2, TM-Shc1 and TM-Shc2 oncoproteins promote morphological transformation and E-cadherin down-regulation in IEC-6 cells. (A) Photographs show the typical morphology of IEC-6 cells expressing, or not (Control), the docking-specific TM-Grb2, TM-Shc1 or TM-Shc2, or the oncogenic Met receptor, Tpr-Met. Expression and phosphorylation levels of these oncoproteins in IEC-6 cell lysates were assessed by immunoblot (IB) analyses following immunoprecipitation (IP) with a Met antibody. (B) E-cadherin protein levels were determined by IB analyses of total cell lysate (TCL) prepared from the indicated serum-starved cells. Tubulin protein levels provide a loading control. (C) E-cadherin mRNA expression (Cdh1) was evaluated by semi-quantitative RT-PCR assays performed with total RNA prepared from the indicated serum-starved cells. The analyses were carried out with two sets of primers designed to amplify distinct regions of the rat Cdh1 mRNA. The mRNA encoding for the S18 ribosomal protein is shown as a loading control. Parallel PCR reactions without the RT enzyme indicate that the amplified products did not arise from genomic DNA contamination. (D) Relative mRNA expression levels of Cdh1 were analyzed by quantitative real-time RT-PCR. These assays were carried out with the same RNA samples and sets of Cdh1 primers. The bar graph shows the mean ± S.E.M. fold-change of Cdh1 mRNA levels relative to Control-IEC-6 cells. The values are from three independent sample sets run in duplicate, normalized to TATA-binding protein (TBP), pumilio RNA-binding family member 1 (Pum1) and ribosomal protein L19 (Rpl19) expression. The primer sequences are listed in Additional file 2.