A: PC3-β1-shRNA or PC3-cont-shRNA transfectants were serum starved and stimulated with or without IGF-1. Cell lysates were immunoprecipitated using Abs to IGF-IRβ and immunoblotted using Abs to phosphotyrosine (p-Tyr, PY20) or IGF-IRβ (left panels). In some experiments, cell lysates from IGF-1 treated PC3-β1-shRNA or PC3-cont-shRNA transfectants were immunoblotted using Abs to p-AKT or AKT (right panels). B: PC3-vector, PC3-β1-shRNA or PC3-cont-shRNA were detached, lysed and immunoblotted using Abs to GLI1, β1 (C-18) or FAK, as a loading control. C: PC3 parental, PC3-vector, PC3-cont-shRNA or PC3-β1-shRNA cells were transfected with pCMV-β-galactosidase and 8×3'GLI-BS pδ51LucII reporter construct. GLI reporter activity was analyzed by luciferase assay. The data were normalized using β-galactosidase activity. Data are expressed as means ±SEM (*p=0.0072).