Figure 3.

Serum deprivation of PASMC for >24 h enables thrombin‐dependent phosphorylation of Akt. PASMC were serum deprived for 48 h, thrombin (A) or PDGF (B) was then prepared with the conditioned (cond.) media that the cells had been grown in or with fresh media and added to the cells. Western blotting for total and phospho‐Akt was then performed. Representative Western blots show thrombin‐mediated phosphorylation and total protein of Akt (C) and ERK (D) in either serum‐deprived media (0.1% FBS) or in serum‐containing cell growth media (10% FBS). (E) shows the time dependence of the phosphorylation of Akt in serum‐starved cells with PDGF‐ or thrombin‐ treated for 15 min after serum starvation. Time course of p‐Akt in PDGF‐ and thrombin‐treated groups shown in arbitrary units. All blots were representative of 3 independent sets of experiments. **P <0.01 versus 0 h.