miR-136 Modulates TGF-β1-Induced Proliferation Arrest by Targeting PPP2R2A in Keratinocytes

Figure 3

PPP2R2A was a direct target of miR-136. (a) There were three potential miR-136 binding sites in PPP2R2A 3^′-UTR based on the TargetScan database; the conservation of the miR-136 binding seed regions among different species was shown in shading and mutations were shown in italics. Fragments containing wild-type (wt) or mutant (mut) miR-136 binding sites in human PPP2R2A 3^′-UTR were cloned downstream of the luciferase reporter gene separately. ((b)–(d)) Luciferase reporter assay ( for each group). Cos-7 cells were cotransfected with a 3^′-UTR reporter construct and the miR-136 mimics or miR-NC, and the results showed that site 1 and site 2 were the direct targets of miR-136. Luciferase activity/renilla activity was applied as the baseline control for the experiments using the same reporter. Data represent mean ± SD. . (e) Western blot analyses of PPP2R2A expression in HaCaT cells transfected with miR-136 mimics or miR-NC. GAPDH was used as loading control.

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