Figure 4

Effects of TIG1A and TIG1B on GRK5 and TCF7 expression in HCT116 cells. (A) Real time RT-PCR analysis of gene expression in inducible stable cells. Stable HCT116 clones were treated with 5 nM MFP for 24 h. Total RNA was extracted and relative levels of the indicated mRNAs were measured by real-time RT-PCR. Data represent the mean and SD of fold changes in gene expression performed in triplicate after normalisation to the results obtained from vehicle-treated control stable cells. Results shown are representative of 2 independent experiments. (B) Western blot analysis of the effects of TIG1 isoforms on GRK5 and TCF7 expression in stable cells. Stable HCT116 clones were treated with 5 nM MFP for 0 to 48 h. Cell lysates were prepared, and expression of TIG1A, TIG1B, GRK5, TCF7 and β-actin was determined by Western blot analysis. (C) Up-regulation of GRK5 expression in HCT116 cells transiently transfected with a TIG1A expression plasmid. Cells were transiently transfected with constitutive TIG1A expression vector for 0 to 72 h. TIG1A, GRK5 and β-actin expression from total cellular lysates was determined by Western blot analysis. Ctrl, control.