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. 2015 Mar 8;6(10):7774–7787. doi: 10.18632/oncotarget.3484

Figure 5. MiR-200c directly targets the CYP1B1 3′-UTR.

Figure 5

(A) Sequence and location of MCS1 and 2 in the 3′-UTR of human CYP1B1 mRNA. (B) Diagram of reporter constructs containing the wild type or MCS-deleted 3′-UTR of human CYP1B1. (C) Relative luciferase activity after transfection of reporter constructs containing the wild type or MCS-deleted 3′-UTR of CYP1B1. **P < 0.01; ***P < 0.001; n.s.: non-significant (D) Relative luciferase activity after co-transfection of reporter constructs containing the wild type or MCS-deleted 3′-UTR of CYP1B1with either miR-200c inhibitor or precursor. *P < 0.05; **P < 0.01; n.s.: non-significant (E-G) After miR-200c inhibitor transfection, relative miR-200c expression was analyzed by RT-PCR (E), CYP1B1 protein level was determined by Western blot (F) and enzyme activity was measured with P450-Glo assay (G) in 786-O cells. *P < 0.05; ***P < 0.001 (H-J) After miR-200c precursor transfection, relative miR-200c expression was analyzed by RT-PCR (H), CYP1B1 protein level was determined by Western blot (I) and enzyme activity was measured with P450-Glo assay (J) in A498 cells. **P < 0.01.