Fig. 5.

Snail and Slug Mediate the Regulation of E-Cadherin and Cell Migration by E2

A, BG-1 cells were transfected with either 250 nm of Snail siRNA (sequence no. 1), Slug siRNA (sequence no. 1) or nonspecific siRNA (NS siRNA) for 48 h. Efficacy of the siRNAs was determined by Western blot analysis. B, Eight hours after transfection with the siRNAs, BG-1 cells were treated with vehicle (Cont) or 10⁻⁷ m E2 for an additional 48 h. Whole-cell lysates was extracted and E-cadherin protein (E-Cad) was detected by Western blot. β-Actin was used as a loading control. C, BG-1 cells were transfected with the human E-cadherin promoter luciferase construct and pSV-βGal in combination with either one of the siRNAs. Eight hours after transfection, cells were incubated with vehicle (Cont) or 10⁻⁷ m E2 for an additional 48 h. The luciferase activities were calculated relative to the promoterless vector (pGL3-Basic) and expressed as fold change relative to vehicle control. The data are shown as mean ± sd of three repeated experiments. D, Scratch was made by scraping the monolayer 24 h after transfection of the siRNAs, and cells were incubated with medium containing vehicle (Cont) or 10⁻⁷ m E2. Wound closure was photographed after 24 h of cell migration. The amount of wound repair was expressed as uncovered area at the indicated time compared with initial uncovered area of vehicle-treated control at time zero. Values are the mean ± sd of three separate experiments. *, P < 0.05 compared with control. Cont, Control.