Figure 5

Effect of Pdp1 overexpression on cardiomyogenesis in differentiating EBs. R1 mESCs were transduced with empty vector or Pdp1 (mock or PDP1 groups). (a) Changes in Pdp1 mRNA expression during differentiation were confirmed using quantitative real-time PCR. (b) PDH complex activity was analyzed in all groups. (c) PDP1 and the PDH complex protein PDH E1α expression was evaluated by western blotting with specific antibodies. (d) After Pdp1 gene overexpression, cardiac muscle-specific molecular markers (α-actinin, α-MHC, cTnI, cTnT, Cx43, Flk1, Mef2c, MLC2) were analyzed by quantitative real-time PCR in Mock and PDP1 group (D8 mock vs D8 PDP1, D24 mock vs D24 PDP1). (e) The beating rates of EBs and (f) BPM were measured during differentiation. (g) Both groups were immunostained using the cardiac-specific antibody cTnT. (h) Cardiomyocyte marker α-actinin expression was analyzed by western blotting in D0 and D8 mock and Pdp1 overexpressed cells. *P<0.05, ^#P<0.01, ^†P<0.001 vs mock. BPM, beat-per-minute; EB, embryoid body; mESC, mouse embryonic stem cells; mRNA, messenger RNA; PDH, phosphate dehydrogenase.