The 12A10 scFv reacts intracellularly with Pyk2, reduces Pyk2 phosphorylation, and inhibits glioma cell migration. A, whole cell lysates of SF767-GFP control cells or SF767-12A10 scFv cells were immunoblotted with anti-Pyk2 mAb (top). Lysates of SF767 control cells or SF767-12A10 scFv cells were immunoprecipitated with the polyclonal anti-Pyk2 antibody and the precipitates blotted with the anti-Pyk2 mAb (middle). The blots were stripped and reprobed with the anti-HA antibody to show that the 12A10 scFv was associated with endogenous Pyk2 (bottom). B, lysates of SF767-GFP control cells or SF767-12A10 scFv cells were immunoprecipitated with polyclonal anti-Pyk2 antibody and the precipitates immunoblotted with the anti-phosphotyrosine antibody pY20. Blots were stripped and reprobed with anti-Pyk2 antibody to verify equal amounts of Pyk2 in the immunoprecipitate. C, radial migration assay of SF767-GFP and SF767-12A10 scFv cells on 10 µg/mL laminin. *, P < 0.05.