Figure 1.

Inducible knockdown of Zeb1 in PC-3 human prostate cancer cells results in enhanced expression of epithelial proteins. Mesenchymal human PC-3 prostate cancer cells were engineered to knockdown expression of the master epithelial-to-mesenchymal (EMT) regulator Zeb1 using the SMARTvector inducible lentiviral shRNA system (Dharmacon), which features Tet-on^® induction of the target shRNA in the presence of doxycycline (Dox). (A,B) Immunoblot analysis of Zeb1 protein expression in the presence or absence of Dox (72 h) in Zeb1^KD (Zeb1 knockdown), control (ctrl) PC-3 cells, or LNCaP cells. (C,D) Immunoblot analysis of E-Cadherin, EpCAM, Vimentin and N-cadherin in Zeb1^KD or ctrl cells 72 h after Dox induction. Representative immunoblots are shown and amido black staining of total protein was used as a loading control. Quantitative data is presented as mean ± standard error of the mean (SEM) fold-change in expression relative to ctrl cells (n = 3). α = significantly different than ctrl no Dox. β = significantly different than ctrl with Dox. δ = significantly different than Zeb1^KD no Dox. γ = significantly different than PC-3 parental ε = significantly different than LNCaP (p ≤ 0.05).