Adhesion-dependent MLEC signaling. (A) Serum-starved MLECs were detached, suspended for 1 hour, and plated on Matrigel-coated plates for the indicated times. Equal amounts of lysate were probed with antibodies to activated Akt (p-Akt), activated Raf (p-Raf), and activated eNOS (p-eNOS). Numbers represent p-eNOS (from densitometry) normalized for eNOS amounts. (B) Serum-starved MLECs were plated on Matrigel and analyzed for Rac and Cdc42 activation at the indicated times. Western blots in this figure are each representative of 3 experiments with different cell preparations.