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. 2009 Dec 29;107(1):264–269. doi: 10.1073/pnas.0907904107

Fig. 5.

Fig. 5.

DDIT4 inhibition by miR-221. (A) Luciferase activity 72 h after cotransfection of 293T cells with psiCHECK-2-UTRs vectors, premiR-221, premiR negative control (NC), and LNA antimiR-221. Data are means ± SD of four independent experiments performed in triplicate. Renilla luciferase activity was significantly decreased for 3′ UTRs of positive controls (Kit, p27, CDKN1C) and for DDIT4, CREBZF, MYBL1, TBK1, DKK2, and BNIP3L (P < 0.05, Student’s t test). (B) Luciferase activity at 72 h in 293T cells cotransfected with premiR negative control (NC, black) or premiR-221 (gray) and luciferase reporters containing the wild-type (WT) or mutated miR-221 binding sites (mS1 and mS2) of human DDIT4 3′UTR or vector (V) (*, P < 0.01, Student’s t test). (C) 293T, Huh6, and NU.K.-1 cells were transfected with vector expressing miR-221, miR-222, or empty vector, and protein levels were checked by Western blotting with the indicated antibodies. One representative of three independent experiments is shown.