Figure 4.

The promigratory effect of PDK1 requires its PH domain and its catalytic activity. (A) ECs were infected with retroviruses carrying PDK1 with a deleted PH domain (ΔPH), PDK1 with of the first 50 amino acids deleted (Δ50), PDK1 kinase-dead (KD), PDK1 mutated in the PIF pocket (L155E), and retroviruses not carrying anything (vector); these cells were used in chemotaxis assays in presence of a gradient of VEGF-A (10 ng/ml). Migration index is calculated assigning a value of 1 to the number of vector cells migrated in the absence of stimulus; data were plotted as the mean ± the SD of six independent experiments. Statistical significance (*, P < 0.01) is shown for VEGF-A–stimulated EC-PDK1-Δ50 and PDK1-L155E compared with vector ECs. (B) Cells were serum deprived for 2 h, and then stimulated or not stimulated with VEGF-A for 10 min; 50 μg of each lysate was immunoblotted with the indicated antibody. Western blots shown are representative of five experiments performed with similar results.