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. Author manuscript; available in PMC: 2016 Oct 23.
Published in final edited form as: Circ Res. 2015 Sep 2;117(10):891–904. doi: 10.1161/CIRCRESAHA.115.306624

Figure 1. The abundance of miR-206 in CMs is regulated by YAP.

Figure 1

(A and B) Neonatal rat CMs (NRCMs) were transduced with Ad-LacZ or Ad-YAP. In A, Northern blot analyses were conducted using miR-206 and U6 probes, and the results were quantified by densitometry. In B, qRT-PCR analyses were conducted with miR-1, miR-206 and 5S rRNA primers. N=3. (C) CMs were transduced with either Ad-LacZ or Ad-Mst1 and Northern blot analyses were conducted using a miR-206 probe (left panel) or western blot was performed to determine phosphorylation status of YAP (right panel). N=3. (D) A schematic representation showing the location of E-boxes and ChIP primers in the miR-206 enhancer (upper). The wild-type and mutant E-box sequences are shown (lower left). Nine wild-type or mutated E-box sites were included in the promoters of the luciferase reporter used in (E) and (F) (lower right). (E and F) CMs were transduced with Ad-YAP, Ad-Mst1 or Ad-LacZ and then transfected with luciferase reporter constructs harboring either a control, wild-type E-box or mutated E-box sequence in the promoter. N=4 (E) and 3 (F). (G) YAP-overexpressing CMs were harvested for chromatin immunoprecipitation (ChIP) assays. ChIP assays were conducted with antibodies against YAP or a control immunoglobulin G (IgG) and specific Ebox primers. PCR using input DNA as a template served as an internal control. N=3.