1. Fig. 4. miR302-367 promotes cardiomyocyte proliferation through regulation of Hippo pathway kinases.
miR promotes cardiomyocyte proliferation through regulation of Hippo pathway kinases. (A) Expression of Mob1b, Lats2, and Mst1 in Nkx2.5cre:R26R-miR Tg/+ mutant hearts at E18.5 and Nkx2.5cre:R26R-miR flox/flox null mutant hearts at E12.5 by qRT-PCR. (B) Luciferase reporter assays showing that miR can repress Mst1, Lats2, and Mob1b expression through their respective 3′UTRs. This repression can be reversed by mutations of the miR binding sites. (C) Confocal fluorescence microscopy of phospho-Yap and nuclear staining of Yap, with or without DAPI, in ventricular cardiomyocytes of Nkx2.5cre:R26R-miR Tg/+ mouse hearts at E18.5. Cytoplasmic and nuclear ratio for total Yap protein was quantified using Fiji software. (A to C) *P < 0.05, **P < 0.01 versus Nkx2.5cre control hearts (Student’s t test). (D) Overexpression of miR in primary mouse neonatal cardiomyocytes. Cardiomyocyte proliferation was quantified using Ki67 immunostaining. **P < 0.01 versus Yap shRNA control. Scale bars, 100 μm. (E) Proposed model of miR promoting cardiomyocyte proliferation through regulation of Hippo pathway kinases. (A to D) Data are means ± SEM (n = 3 for A, C, and D; n = 5 for B).
Ying Tian et al., Sci Transl Med 2015;7:279ra38
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