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. 2018 Jan 29;22(4):2162–2176. doi: 10.1111/jcmm.13490

Figure 3.

Figure 3

KDM6B expression is specifically regulated by miR‐99a. (A) miRNA target prediction using a combination of the following computational algorithms: TargetScan, microRNA.org and miRBase. (B) Schema and sequence of predicted miR‐99a binding site on wild‐type (WT) KDM6B UTR, sequence of miR‐99a and sequence of mutated (MUT) KDM6B UTR. (C) Wild‐type and mutant KDM6B 3′UTR were, respectively, inserted into a pmirGLO Luciferase Vector, which were transfected into C3H10T‐1/2 cells. Cells were cotransfected with either control or miR‐99a mimic/inhibitor oligomers. miR‐99a effectively suppressed luciferase reporter activity, while the mutations significantly impaired the activity of miR‐99a, relieving repression of luciferase activity. (D) qRT‐PCR analysis of the relative KDM6B mRNA expression levels in C3H10T‐1/2 cells. There was no change of KDM6B mRNA expression after miR‐99a mimics transfection. (E) Western blotting showed that KDM6B protein level was all significantly down‐regulated by miR‐99a mimics treatment. Data are shown as mean ± S.D. *P < 0.05.