Shear-induced αvβ3–Shc association and JNK
activation were inhibited when HUVECs were seeded on slides coated with
LM609 anti-αvβ3 mAb. The slides for lanes 1
and 2 were coated with FN (5 μg/cm2) and those for
lanes 3 and 4 were coated with FG (5 μg/cm2). The
slides for the remaining lanes were first coated with an anti-mouse IgG
followed by LM609. HUVECs were seeded on the slides and allowed to
adhere for 2 h (lanes 1–6) or 30 min (lane 7). (A)
The seeded HUVECs were kept under static condition (C) or subjected to
shear stress (S). Lane 7 (Ad) served as a positive adhesion control to
show that αvβ3–Shc association occurred in
static HUVECs at 30 min after onset of adhesion, but not at 2 h
(lane 6). The cell lysates were subjected to IP with LM609, followed by
IB with an anti-Shc pAb. (B) HUVECs were kept as static
controls (C), subjected to shear stress (S), or exposed to UV
irradiation of 80 J/m2 for 5 min (UV, lane 8). The
cells were lysed and JNK kinase activity was assayed by using GST-c-Jun
as the substrate. Gels are representative of three separate
experiments.