Figure 3.

Analysis of expressions of HC-related markers in Math1-ES-derived EB outgrowths. (a) Analysis of expressions of Math1, as well as myosin6, myosin7a, Brn3c, and α9AchR as HC-related markers in EBs cultured with or without Dox for 2 weeks. (b) Immunocytochemical analyses of myosin6 (My6), Brn3c, and α9AchR in EB outgrowths cultured in the presence of Dox for 2 weeks. Bar=50 μm. (c) Frequencies of immunopositive cells for My6, Brn3c, and α9AchR in EB outgrowths cultured for 2 weeks with or without Dox (n=3). Without Dox, immunopositivity for myosin6, Brn3c, and α9AchR was seen at very low levels. In contrast, a remarkably increased number of immunopositive cells was detected in EB outgrowths cultured with Dox. (d) Simultaneous expressions of My6, Brn3c, and α9AchR in EB outgrowths cultured with or without Dox (n=3). In cultures with Dox, the three markers were simultaneously expressed at high frequencies. GAPDH, glyceraldehyde 3-phosphate dehydrogenase; DAPI, 4,6-diamidino-2-phenylindole