Figure 3

EPA treatment of HT29 cells leads to significant changes of EphB4 and Vav2 protein expression. EPA treatment of HT29 cells leads to significant changes of EphB4 and Vav2 protein expression levels as determined by western blotting. Densitometry measurements were taken on a Bio-Rad multi-imager and all data is expressed relative to the house-keeper protein, β-actin, allowing direct comparison of protein expression data to gene expression data as shown in figure 2. EPA treated cells are represented by black bars, AA treated cells by grey bars and untreated cells by white bars. Data are presented as the mean and SD of n = 3 (n = 2 at 4 hr). A) EphB4 protein was not detectable until 8 hr after media change. At 8 hr EPA treatment led to lower protein levels compared to both untreated and AA treated HT29 cells (p = 0.007 and p = 0.034 respectively). There was a highly significant increase in protein levels between 8 hr and 24 hr in all treatment groups (p < 0.005). B) Vav2 protein expression was significantly up-regulated after 4 hr irrespective of treatment (p < 0.05). At 8 hr levels of Vav2 protein were significantly elevated by EPA treatment as compared to untreated control cells (p = 0.043).