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. Author manuscript; available in PMC: 2010 Aug 1.
Published in final edited form as: Hepatology. 2009 Aug;50(2):472–480. doi: 10.1002/hep.22989

Figure 5. CDX2, GATA6 and NLK as direct targets of miR-181s.

Figure 5

(A) Predicted duplex formation between the 3’-UTR sequences of human CDX2, GATA6 and NLK and miR-181b. The boxes highlight the seed sequences in miR-181b. The underlined bases were mutated to UUACA in mutant 3’-UTR plasmids. (B, C) The expression levels of CDX2, GATA6 and NLK in Hep3B cells transfected with miR-181 inhibitors (B) or in HuH1 cells transfected with miRVec-181s (C), as determined by western blotting. (D) Luciferase activities of various reporter plasmids in HuH1 cells co-transfected with miRVec-control, 181a, 181b, or 181c/d. Each experiment was performed in triplicate and the luciferase activity was shown as mean ± SD. A student t-test was employed. * refers to p<0.01. Levels of GATA6 (E) or NLK (F) in HuH1 cells transfected with control, GATA6 siRNA or NLK siRNA were determined by Western blotting (left panel) or qRT-PCR (middle panel). The Relative expression ratio (GATA6 siRNA or NLK siRNA versus control siRNA) of UGT2B7, CYP3A4, CCND1 and TACSTD1 genes in HuH1 cells is shown as mean±SD in log(2) scale (right panel).