Figure 3.

Immunocytochemical analysis of specific markers for HCs in EB outgrowths. (a) After 2 weeks of differentiation of EBs in ES-M or ST2-CM, EB outgrowths were analyzed for HC-related markers (Brn3c, myosin7a, calretinin, Math1, myosin6, and α9AchR) using an immunohistochemical method. ST2-CM dramatically induced the emergence of immuno-positive cells for those markers in EB outgrowths. Scale bar=20 μm. (b) Simultaneous expressions of Brn3c, myosin6, and calretinin with Math1 were examined in EB outgrowths cultured with ST2-CM using an immunocytochemical method. Scale bar=10 μm