Emodin blocks TGF-β-induced EMT in BT20 cells.

<p>(A) BT20 cells were treated with various concentrations (1–10 ng/mL) of TGF-β for 24 h. The cells were then harvested and lysed for the detection of E-cadherin, vimentin, and β-actin. (B) BT20 cells were pretreated with DMSO (control) or 20 μM emodin for indicated time periods and then stimulated with TGF-β (2 ng/mL) for 24 h. The cells were harvested and lysed for the detection of E-cadherin, vimentin, and β-actin. (C) BT20 cells were pretreated with DMSO (control) or increasing emodin concentrations (5–20 μM) for 2 h and then stimulated with TGF-β (2 ng/mL) for 24 h. The cells were harvested and lysed for the detection of E-cadherin, vimentin, and β-actin. Western blot data presented are representative of those obtained in at least 3 separate experiments. (D) For wound healing assay, confluent BT20 monolayer was pretreated with DMSO (control) or increasing emodin concentrations (5–20 μM) for 2 h, cells were scratched by pipette tips and washed to remove the debris and following by fresh medium containing 0.5% serum with emodin. Cells were then incubated with 2 ng/mL TGF-β for 24 h. TGF-β-induced cell motility was determined by measuring the closure of wound. Data were plotted by mean ± S.D. (<i>n</i> = 3). The closure distance of the control cells was set to 1. Emodin significantly inhibited TGF-β-induced cell motility (*, <i>p</i> < 0.05; **, <i>p</i> < 0.01).