Figure 5
Transient transfection of HSA/c with NGF-siRNA. (A) NGF-siRNA significantly reduced NGF secretion from HSA/c by 85% in ELISA (*P<0.0001). Twenty-four hours after transient transfection of NGF-siRNA, medium was changed to serum-free medium. At 72 h after transfection, conditioned media were collected. NSC: nonspecific control RNA. (B) Western blot analysis for NGF, TrkA, and phospho-TrkA. After transfection with siRNA, the NGF expression decreased to the level nearly undetectable. Western blot also revealed reduction of phosphorylated TrkA. There was also reduced expression of TrkA after siRNA transfection, but its reduction rate is less than that of phospho-TrkA, as clearly shown by NGF-siRNA/NSC expression ratio. (C) Transwell migration assay. Nerve growth factor-siRNA significantly decreased cell migration (*P=0.0009).
