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Fig. 2. (i) {alpha}8ß1 mediates spreading on LAP-TGFß1. {alpha}8-transfected or mock-transfected AtT20 cells were plated on 5 µg/ml of recombinant LAP-TGFß1 in serum-free media for 24 or 48 hours. At 24 hours, mock transfected AtT20 cells remain rounded and unattached on LAP-TGFß1 (A), whereas {alpha}8-transfected cells attach and spread on LAP-TGFß1 (B). Further spreading is seen at 48 hours (C). (ii) Adhesion of {alpha}8ß1 to LAP-TGFß1 results in FAK phosphorylation. {alpha}8-transfected or mock-transfected AtT20 cells were plated on 0.01% poly-L-lysine (PLL), 5 µg/ml fibronectin (FN), 5 µg/ml LAP-TGFß1 (LAP1) or 5 µg/ml RGE-LAP-TGFß1 (RGE) for 30 minutes in serum-free media. Cells were lysed in buffer containing phosphatase inhibitors, immunoprecipitated with anti-FAK antibody followed by western blotting with either anti-phosphotyrosine antibody PY20 (top panel) or anti-FAK antibody (bottom panel).