Fig. 2.

E2A deficiency altered cardiac-specific differentiation of hESCs.

(a) Representative western blots showing E2A levels in H7 and H9 knockout homozygous clones. (b) Expression pattern of cardiogenic mesoderm markers (MESP1); cardiac progenitor cells markers (NKX2-5, ISL1) and cardiomyocytes structural genes (TNNT2 and MYL7, except for MYH6) were down-regulated in E2A knockout differentiated cells. Data are shown as the mean ± SEM from at least three independent experiments. * P value < 0.05, ** P value < 0.01, *** P value < 0.001, # P value < 0.0001(Two-tailed Student's t-test). (c) Quantification of cTnT positive cardiomyocytes in E2A knockout CMs by flow cytometry. Gray shapes represented the blank control. (d) IF analysis of differentiated cardiomyocytes generated from H7 and H9 hESCs, showing attenuated cTnT relative intensity. White arrows pointed at small size CMs. (e) Quantitation of cTnT intensity was weakened in E2A knockout CMs. (f) Quantitation of CMs cell area was reduced in E2A knockout CMs. (g) Contraction traces of wildtype and E2A knockout CMs. (h) Rising beating rates were observed in CMs derived from E2A knockout CMs. (i) E2A knockout CMs showed attenuated contractile force, n >100.* P value < 0.05,** P value < 0.01, *** P value < 0.01, # P value < 0.0001(Two-tailed Student's t-test).