Figure 5.

Involvement of integrin related molecules in response to collagen I. (A) NMuMG/E10 cells were extracted 3 h after plating on noncoated, collagen I-coated, or fibronectin-coated dishes. RIPA extracts were immunoblotted for phospho-FAK (Tyr 577) and total FAK. (B) Lysates (300 μg of protein) were immunoprecipitated with anti-paxillin mAb and resolved by SDS-PAGE. Phospho-tyrosine and total paxillin were detected by PY20 mAb and paxillin mAb immunoblots, respectively. (C) NMuMG/E10 cells infected with shEGFP (a and b) or shIntegrin β1 (c and d) were cultured on noncoated (a and c) or collagen I-coated (b and d) dishes. Two days after seeding, phase pictures were taken using a 10× objective. (D) Two days after seeding on noncoated (non) and collagen I-coated (col) dishes, cells were extracted, and 30 μg of protein was resolved by SDS-PAGE and immunoblotted for integrin β1, N-cadherin, and tubulin.