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. Author manuscript; available in PMC: 2013 Jan 1.
Published in final edited form as: J Cell Mol Med. 2012 Jan;16(1):160–173. doi: 10.1111/j.1582-4934.2011.01282.x

Figure 8. Regulation of miR-181 expression by Twist in HCC stem cells.

Figure 8

Panel A: The schematic diagrams of miR-181 genes showing the Twist1 binding e-boxes are located in 5’-promoter regions of miR-181a-1/miR-181b-1. Panel B: Real-time PCR confirmed reduced expression of mature miR-181a in HSC-SR and HSC-DF cells in Twist silencing group when compared with expression in their respective controls. Panel C&D: Total RNAs were isolated using Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit and real-time PCR assay was carried out using Taqman MicroRNA Assay kit. Increased miR-181a and miR-181b binding with Twist were observed in HCC cancer stem cells under self renewal or differentiation condition. * p < 0.05 when compared with respective controls. Panel E&F: Overexpression of miR-181a and miR-181b inhibited RASSF1A, TIMP3 and NLK protein expressions in HSCs. Representative immunoblots (Panel E) and quantitative data (Panel F, mean ± se) from 4 separate blots are shown. *p < 0.05 relative to expression in controls.