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. 2014 Jan 14;110(5):1260–1268. doi: 10.1038/bjc.2013.829

Figure 6.

Figure 6

FOXO1 and p27Kip1 are direct targets of miR-196a. (A) Expression of p21Cip1 and cyclin D1 in C33A and CaSKi cells after transfection with miR-196a or NC by western blot analysis. α-Tubulin was used as a loading control. (B) Expression of p21Cip1and cyclin D1 in indicated cells by real-time PCR. (C) Predicted miR-196a target sequences in the 3'-UTRs of FOXO1 and p21Cip1 genes. (D) Expression of p27Kip1 and FOXO1 in indicated cells by western blot analysis. α-Tubulin was used as a loading control. (E) Relative FOXO1 reporter activities in the indicated cell lines. (F) Luciferase assays on C33A and CaSKi cells transfected with the pGL3 control reporter, pGL3-target-3'-UTR reporter, or pGL3- target -3'-UTR-mut reporter and increasing amounts of miR-196a mimic or miR-196a inhibitor oligonucleotides, as indicated. Error bars represent mean±s.d. from three independent experiments. *P<0.05.