Effect of ectopic twist expression. CMV-driven eukaryotic expression constructs for CAT (control) and twist were introduced into PLC, HepG2, and Huh7 cells using Superfect™ chemical transfection reagent. A. After a 48-h incubation period, cells were lysed. Equal amounts of protein were analyzed by SDS-PAGE and immunoblotted with anti-twist and anti-α-actin antibodies. Shown are representative blots of more than three independent studies. B, C. After a 24-h incubation period, cells were quiesced for 24 h in DME with 0.1% dialyzed FBS. Then cells were cultured with 10% FBS. Cell proliferation (B) and cell migration activities (C) were assessed as described in Materials and Method. All results are expressed as a ratio to control CAT cDNA transfected PLC (CAT; clear bars, twist; black bars). The data are the 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 CAT cDNA transfected cells).