FIG. 5.

Efficacies and specificities of intracellular signaling inhibitors. Rat PA SMCs were maintained in complete medium and transferred to DMEM containing 0.2% FCS 24 h before treatments. Cells were treated with 20 μM U0126, 40 μM SP600125, 40 μM LY294002, 10 μM Akt inhibitor, or 40 nM rapamycin for 30 min. “No Add'n” indicates cells not treated with an inhibitor. Duplicate wells either were left untreated or were subjected to the addition of PDGF to a final concentration of 25 ng/ml. After 30 min of incubation, the cells were lysed, and 25 μg of lysate protein was resolved on 10% polyacrylamide-SDS gels and transferred to PVDF membranes. The blots were subjected to Western blot analysis with antibodies to the phosphorylated and total signaling intermediates indicated to the left of each panel.