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Fig. 3 | Stem Cell Research & Therapy

Fig. 3

From: Hsa-miR-335 regulates cardiac mesoderm and progenitor cell differentiation

Fig. 3

MiR-335 regulates earlier stages of hESC cardiac differentiation. a Immunocytochemistry analysis showed that overexpression of hsa-miR335-3p or hsa-miR335-5p on day 0 did not affect the final outcome of cardiac differentiation on day 12 (cardiomyocytes), as detected by MYH6 expression in all groups. b Overexpression of both hsa-miR335-3p (top) and hsa-miR335-5p (bottom) resulted in significant elevation of BRACHYURY (mesoderm marker) expression. Consistently, hsa-miR335-3p downregulation resulted in reduced expression of BRACHYURY. Western blot results indicate an increased level of BRACHYURY following the mimic-3p and mimic-5p treatment, compared to mimic-scr control (right). c No significant expression alteration was detected for MESP1 following alterations in hsa-miR335 expression. d, e Expressions of GATA4 and NKX2-5 (main cardiac progenitor markers) were both increased following miR-335-3p (top) and miR-335-5p (bottom) overexpression. Downregulation of miR-335 had reverse effect on both of these cardiac progenitor markers. f Expression of HCN4 (the first heart field marker) was significantly increased following miR-335-3p (top) and miR-335-5p (bottom) overexpression. g Alterations in the expression of miR-335-3p (top) and miR-335-5p (bottom) had a significant reverse effect on ISL1 (the second heart field marker) expression. h, i Has-miR-335-3p (top) overexpression increased CNX43 expression but decreased TNNT2 expression. However, has-miR-335-5p decreased CNX43 but enhanced TNNT2 expressions. RT-qPCR data are presented as mean ± SEM normalized against mimic-scr and siRNA-scr. GAPDH was used as a housekeeping gene

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