Phosphorylation of Akt via the CCR9/TECK interaction.
(A) At different time points, 1 × 107 MOLT4 cells were serum starved and stimulated with 50 nM TECK. Phosphorylated Akt was detected by means of phospho-specific Akt antibody (S473). Subsequently, the used blot was stripped and reprobed with pan-Akt antibody. For inhibitor experiment, the serum-starved MOLT4 cells were pretreated with WM or PT for 1 hour and stimulated with 50 nM TECK for 30 minutes. Western blot analysis was conducted by means of phospho-specific Akt antibody (S473). Positive control is cell lysate prepared from NIH3T3 cells stimulated with platelet-derived growth factor (PDGF), whereas negative control is cell lysate prepared from unstimulated NIH3T3 cells. (B) Cos7 cells were transiently transfected with CCR9 alone or with CCR9 and Akt coexpression vector and stimulated with 50 nM TECK. Phosphorylated Akt or total Akt was detected as described above. The bottom pictures in each panel represent expression levels of total Akt. These results are 1 of at least 3 reproducible experiments.
