Differentiation of an αMHC-GFP reporter hESC line recapitulates cardiac myogenesis and identifies developmentally regulated miRNAs.

<p>mRNA expression profiles of hESC-derived, αMHC-GFP⁺ myocardial precursors sorted at days 8 (<b>A, C, E</b>) and 14 (<b>B, D, F</b>) of differentiation demonstrated upregulated expression (red) of genes involved in cardiac myogenesis (<b>A, B</b>), cellular differentiation into CMs (<b>C, D</b>), and withdrawal from the cell cycle (<b>E, F</b>). Abl1, c-abl non-receptor tyrosine kinase; ALK, BMP receptor; ATF2, activating transcription factor 2; ATM/ATR, ; ATP2A2, cardiac calcium transporting ATPase; BMP, bone morphogenic protein; BMPR, BMP receptor; CDK, cyclin-dependent kinase; DP-1, transcription factor Dp-1; E2F, E2F transcription factor; EBP1, ErbB3 binding protein; GATA, GATA binding protein; HAND, heart and neural crest derivatives expressed factor; HDAC, histone deacetyltransferase; ISL1, islet-1; MEF2C, myocyte enhancer factor 2C; MLC2v, ventricular myosin light chain 2; MYH6, α-myosin heavy chain; MYL3, ventricular myosin light chain 3; MYL4, atrial myosin light chain 4; MYLPF, fast skeletal myosin light chain; MYOCD, myocardin; MYOM1, myomesin 1; NKX2.5, NK2 transcription factor related locus 5; NPPA, natriuretic peptide A; NPPB, natriuretic peptide B; NR2F, nuclear receptor subfamily 2, group F; p15INK4, cyclin-dependent kinase inhibitor 2B; p16INK4, cyclin-dependent kinase inhibitor 2A; p21Cip1, cyclin-dependent kinase inhibitor 1A; p27Kip1, cyclin-dependent kinase inhibitor 1B; p53, 53 kD tumor protein ; Rb, retinoblastoma protein; SCF, Skp-Cullin-F-box containing E3 ubiquitin ligase complex; SIN3A, SIN3 transcription factor homolog A; SMAD, SMA/mothers against decapentaplegic; SMYD1, SET and MYND domain containing protein; Suv39H1, suppressor of variegation 3–9 homolog 1; TAK1, mitogen-activated protein kinase kinase kinase 7; TBX, T-box transcription factor; TGF-β, transforming growth factor-β; TnI, cardiac troponin I; TNNC1, troponin C type 1; TNNI1, troponin I type 1; TNNT2, cardiac troponin T ; TTN, titin; WNT11, wingless-type MMTV integration site family member 11; ZFPM2, zing finger protein multitype 2; β-MHC, β-myosin heavy chain. <b>G</b>) Relative miRNA expression in αMHC-GFP⁺ hESCs sorted at days 8 and 14 of differentiation (two samples each), compared to undifferentiated αMHC-GFP reporter hESCs. Heat map shows log₂-fold change (sample intensity/mean intensity for undifferentiated group) for the 156 miRNAs that had statistically significant differences (false discovery rate (FDR)<0.05) of at least 2-fold in any pairwise comparison between the three groups.