Fig. 4.

Immunofluorescence analyses of multilineage ES cell-derivatives using the NC/CNC-specific marker 4E9R. (A) Immunocytochemistry of ES cell-derived non-CNC-related skeletal muscle cells applying 4E9R (green) and the skeletal muscle cell-specific antibody α-Actinin2 (red). (B∼D): Representative immunofluorescence staining of ES cell-derived CNC progeny using 4E9R (in green) and lineage-specific markers as α-Actin2 for smooth muscle cells (B) as well as Nestin (C) and Tubb3 (D) for neuronal cells (all in red). (E∼J) Immunofluorescence analysis of ES cell-derived progeny using 4E9R (in green) and CNC-associated markers such as Mef2c (E), Cx43 (F), Hand2 (G, J), Pax3 (H) and Lbx1 (I) (all in red). White arrows indicate co-expression. Nuclei were counterstained using DAPI (blue). Bars: 50 μm (A∼E, G∼J); 10 μm (F).