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. Author manuscript; available in PMC: 2015 Nov 20.
Published in final edited form as: Biochem J. 2014 Mar 1;458(2):335–341. doi: 10.1042/BJ20130818

Figure 2. NRG-1β treatment from day 0 to 2 of ESC differentiation strongly induces cardiomyogenesis.

Figure 2

(a and b) NRG-1β treatment from day 0 to 2 increases the expression of the cardiac markers Nkx2.5 and Myh6. The qPCR results represent the relative expression normalized to that of PBS vehicle-treated cells at day 0. Measurements were obtained from at least three independent experiments for each time point (*P <0.01). (c) Western blotting showing induction of cTnT protein in NRG-1β-treated mESCs on day 10 in comparison with the PBS-treated controls. An antibody against α-tubulin was used as a loading control. (d) mESCs treated with NRG-1β differentiated to cardiomyocytes that expressed the sarcomere proteins α-actinin (upper panels) and cTnT (lower panels). Confocal images were taken using a Zeiss inverted LSM 510 confocal microscope (× 40). Topro 3, TO-PRO-3™ (Molecular Probes). (e) NRG-1β treatment from day 0 to 2 down-regulated BryT+ at day 3 (*P <0.01), but led to a higher expression of BryT+ at day 4, in comparison with vehicle controls (**P <0.05). The relative RNA expression level of each marker was calculated by normalizing the marker RNA expression at each specific day with the RNA expression at day 0.