Effect of gene silencing using twist siRNA on cell migration. A. After treatment with twist siRNA duplexes, twist protein expression was analyzed by SDS-PAGE and immunoblot using anti-twist antibody. Immunoblotting for α-actin expression levels was used to verify equal loading of cellular protein. Shown are representative blots of more than three independent studies. B. After treatment with twist siRNA duplexes, cell migration assay was performed, as described in Materials and Methods. All results are expressed as a ratio to control siRNA treated HLE (control siRNA; clear bars, twist siRNA; black bars). The data are mean ± SEM of at least three independent studies, each performed in triplicate. Statistical analysis was performed by Student's t-test: *P < 0.01; n.s., not significant (vs control siRNA treated cells).