Figure 6

Roles of intracellular kinases and extracellular stimuli in cTnT expression in SANC cultures. The effects of various chemicals on cTnT expression levels in cell clusters were examined using quantitative RT-PCR after 3 weeks culture. (A) Activation and inhibition of protein kinases A, C and G; the effects of 8-bromo cAMP (500 µM), phenylephrine (3 µM) and 8-bromo cGMP (500 µM; n = 3) on cTnT expression levels are shown in panel a, and those of H89 (10 µM), GF109203 (1 µM) and KT5823 (400 nM; n = 6) are shown in panel b. (B) The roles of CaMK II, ROCK, EGFRK, PI₃K, Akt and GSK3 were examined by analysing the effects of the corresponding inhibitors KN-93 (2 µM), Y-27632 (10 µM), AG1478 (1 µM), LY294002 (10 µM), Akt inhibitor VIII (1 µM) and BIO (3 µM), respectively, (n = 6). (C) Roles of ERK1/2, JNK and p38 were examined by analysing the effects of the inhibitors PD98059 (10 µM), SP600125 (10 µM) and SB203580 (10 µM), respectively, (n = 6). (D) The effects of BMP4 (10 ng/mL), IGF (10 ng/mL), EGF (100 ng/mL), VEGF (50 ng/mL), triiodothyronine (30 nM) and retinoic acid (1 µM; n = 3). Bars denote mean ± S.E.M.; Dunnett’s test; *P < 0.05, **P < 0.01, ***P < 0.001.