Shear-induced integrin–Shc association is ECM-dependent. Glass
slides were coated with FN (5 μg/cm2), CL (6
μg/cm2), LM (5 μg/cm2), or VN
(0.5 μg/cm2). HUVECs were seeded on the slides in
the absence of serum, and exposed to shear stress (12
dyn/cm2) for 30 min or kept as static controls.
(A) The cell lysates were subjected to
immunoprecipitation (IP) with LM609
anti-αvβ3, followed by immunoblotting (IB)
with anti-Shc pAb. Each pair of lanes represents static control (C) and
sheared (S) samples of cells plated on the ECM indicated.
αvβ3–Shc association is demonstrated by
coimmunoprecipitation in the sheared, but not static, HUVECs on FN and
VN. (B) α6β1–Shc
association is demonstrated by coimmunoprecipitation in the sheared
HUVECs on LM only. Gels shown are representatives of three independent
experiments.