Binding of the α_MI domain to Cyr61 and CTGF.

Microtiter wells were coated with the indicated concentrations of Cyr61 (A) or CTGF (B), and blocked with 0.15% PVA. GST-α_MI fusion protein or GST (1 μM each) was added and allowed to bind for 2 hours at 22°C. After washing, bound GST-α_MI or GST was detected using a polyclonal anti-GST antibody followed by an HRP-conjugated secondary antibody. Bound antibodies were detected usingo-phenylenediamine dihydrochloride as the substrate and A₄₉₀ was measured. (C) GST-α_MI was preincubated with vehicle buffer (control), 500 nM YFC118.3 (anti-β₂), or 2LPM19c (anti-α_M) for 30 minutes at 37°C prior to addition to wells coated with 30 μg/mL Cyr61 (open bars) or CTGF (closed bars). Data shown are means of triplicate determinations and error bars represent SEs.