Fig. 4. Expression of 
-SMA is induced in HFL-1 cells by function-blocking antibodies to VN integrins. (A) Protein from HFL-1 cells exposed to P1F6 (anti-vß5), L230 (anti-v), LM609 (anti-vß3) or P5D2 (anti-ß1) was subjected to SDS-PAGE and probed by western blotting using a monoclonal antibody to -SMA. Membranes were stripped and reprobed with a monoclonal antibody for tubulin, as a protein loading control. Blots are representative of at least three separate experiments. (B) HFL-1 cells were incubated with monoclonal antibodies recognizing specific -subunits: L230 (v), P1E6 (2), P3D10H5 (5); specific ß-subunits: 10D5.8 (ß6), P5D2 (ß1), 25E11 (ß3); integrin heterodimers: LM609 (vß3) and P1F6 (vß5) or RGD peptides GRGDdSP and GPenGRGDSPCA for 24 hours. Cells were permeabilised, fixed in methanol and incubated with a monoclonal antibody to -SMA followed by HRP-conjugated rabbit anti-mouse IgGs. Expression of -SMA was then analyzed by ELISA. Results are calculated from triplicate wells of at least four different experiments and are expressed as mean±s.e.m. *Significantly greater -SMA expression compared to control cells, P<0.05.
