Figure 4: Validation of the EMT activity-related expression of microRNAs and proteins. The expression levels of some key mesenchymal-associated microRNAs, that is, mir-200a (microRNA-200a), mir-200b (microRNA-200b), mir-200c (microRNA-200c), mir-141 (microRNA-141), mir-429 (microRNA-429), and mir-205 (microRNA-205), as well as Zeb1 and Zeb2 mRNAs in epidermal cells treated with shikonin for 4 h (a) and 24 h (b) were determined by real-time PCR analyses. Values shown are the mean ± SEM of three experiments. The significance of differences was analyzed by one way ANOVA (; ). (c) Expression of E-cadherin and other mesenchymal-associated markers, MMP2, MMP-9, and vimentin, were measured by Western blot analysis. β-actin was used as gel loading control. Data are representative of triplicate experiments.