Fig. 3
IGF-II overexpression promotes PSC cardiomyogenic differentiation in vitro. a qRT-PCR analysis of IGF-II mRNA levels in 1 × 106 PSC- or ESC-derived cells at different time points after differentiation. Data are presented as the log fold-change in mRNA expression normalized to that in untransfected ESC line D3 cells at day 2 after differentiation. Data are expressed as mean ± SD. *p < 0.05 vs. pME 18s PSCs. n = 8. b qRT-PCR analysis (n = 8) of cardiomyocyte-specific markers (cTnT, α-MHC, and β-MHC) in 1 × 106 PSC- or ESC-derived cells at different time points after differentiation. The mRNA levels for cTnT, α-MHC, and β-MHC were also detected in 1 × 106 cardiomyocytes isolated from mouse fetal (14 days post coitus), neonatal (1-day post-birth), juvenile (2 weeks post-birth), and adult (10 weeks post-birth) hearts, respectively. Data are presented as the log fold-change in mRNA expression normalized to that in ESC line D3 cells at day 10 after differentiation. *p < 0.05 vs. pME 18s ESC- or pME 18s PSC-derived cells at day 10 (D10); #p < 0.05 vs. pME 18s ESC- or pME 18s PSC-derived cells at day 17 (D17); ∆p < 0.05 vs. pME 18s ESC- or pME 18s PSC-derived cells at day 24 (D24), but p > 0.05 vs. juvenile cardiomyocytes. c Western blot analysis was performed to detect the cTnT and cTnI protein expression in cells derived from pME 18s-IGF-II PSCs at different time points after differentiation. β-actin was used as an internal control. d–f To demonstrate the maturation of cardiomyocytes derived at day 24, immunofluorescence study of α-actinin was conducted. d The eGFP fluorescence. e Immunostaining of α-actinin. f Merge of d and e. Scale bar, 25 μm