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. 2001 Jan 23;98(3):1042–1046. doi: 10.1073/pnas.031562998

Figure 4.

Figure 4

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.