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Fig. 5. RGD peptides do not inhibit OPN-induced JB6 cell transformation, however, antibody to OPN significantly reduces OPN-induced colony formation. (A) Cells were pre-incubated with GRGDSP (competitive) or GRGESP (negative control) peptides at 1 or 10 mM for 20 min prior to seeding on top of the 0.5% solidified agar layer with or without 45 µg/ml of OPN and incubated for 14 days. **P< 0.01 significantly different from respective controls, which do not have OPN (designated as OP or P) added. (B) OPN were pre-incubated with molar equivalent of purified anti-OPN or control IgG for 30 min prior to mixing with cells and layered on top of the 0.5% solidified agar. Negative control such as cells incubated with PBS, anti-OPN or IgG were also included. Histogram was plotted as the percent of colony formation relative to OPN-treated cells. The number of clones for the negative controls was in the range of 60 to 35 clones/10 000 cells. *P<0.05 significantly different OPN (OP)-treated cells.
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