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Fig. 2. (i)
8ß1 mediates spreading on LAP-TGFß1.
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
8-transfected cells attach and spread on
LAP-TGFß1 (B). Further spreading is seen at 48 hours (C). (ii) Adhesion
of
8ß1 to LAP-TGFß1 results in FAK phosphorylation.
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).