Fig. 2
Generation of cardiomyocytes from ESC-Rep. a Illustration of cardiomyocyte differentiation protocol of ESCs and stage-specific cell fate as well as representative markers. b Immunofluorescence staining of ESC-Rep-CMs for cardiac structural markers alpha-actinin and troponin T (TNNT2). Nucleus is marked by Hoechst 33342 (blue). Scale bar, 50 μm. Real-time PCR analysis of cardiac-specific gene expression in ESC-Rep-CMs, including MYH6 (c), MYH7 (d), and TNNI3 (e). f Real-time PCR analysis of the three reporter genes (GFP, HSVtk, and Fluc) in ESC-Rep and ESC-Rep-CMs. g Luciferase activity measurement of ESC-Rep and ESC-Rep-CMs. The luciferase activity was normalized to 1 μg protein. h Luciferase activity in ESC-Rep-CMs in 6-well plates. i A robust correlation existed between cell number and bioluminescence signals (R2 = 0.98). All data are presented as the mean ± SEM; Student’s t test; *p < 0.05, and ns, not significant