Fig. 4. miR302-367 promotes cardiomyocyte proliferation through regulation of Hippo pathway kinases. miR302-367 promotes cardiomyocyte proliferation through.

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Fig. 4. miR302-367 promotes cardiomyocyte proliferation through regulation of Hippo pathway kinases. miR302-367 promotes cardiomyocyte proliferation through regulation of Hippo pathway kinases. (A) Expression of Mob1b, Lats2, and Mst1 in Nkx2.5cre:R26R-miR302-367Tg/+ mutant hearts at E18.5 and Nkx2.5cre:R26R-miR302-367flox/flox null mutant hearts at E12.5 by qRT-PCR. (B) Luciferase reporter assays showing that miR302-367 can repress Mst1, Lats2, and Mob1b expression through their respective 3′UTRs. This repression can be reversed by mutations of the miR302-367 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-miR302-367Tg/+ 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 miR302-367 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 miR302-367 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 Copyright © 2015, American Association for the Advancement of Science