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Fig. 2. The interconversion of p105 and p115 HEF1 is regulated by cell adhesion and detachment. (A) A1-F monolayers plated on FN-coated dishes were serum-starved overnight and treated with TGF-ß1 (2.5 ng/ml) for 4 hours to induce HEF1 expression (0 hours). Cells were then detached and suspended in serum-free medium for indicated times. (B) After suspension for 2 hours (indicated as 0 hours of adhesion), A1-F cells were plated onto FN-coated dishes for indicated times. (C) After having been treated with TGF-ß1 (2.5 ng/ml) for 4 hours, A1-F monolayers were detached and suspended in serum-free medium for 1 hour. Suspended cells were then plated onto tissue culture dishes that had been coated with polylysine (PL), fibronectin (FN), collagen (Coll), laminin (LM) or vitronectin (VN) and incubated for 2 hours. Cell lysates were prepared and processed for immunoblotting with anti-HEF1 pAb. Nitrocellulose membranes were stripped and reprobed with anti-p130Cas mAb as loading control.

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