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Fig. 4. Defective embryoid body formation in the Mp–/– ES cells. (A-D) Phase contrast micrographs of Mp+/+ and Mp–/– embryoid bodies. Arrowheads show the lumen formed in the Mp+/+ embryoid body and arrows show the outer layer of cells detaching from the Mp–/– embryoid bodies. (B) Maspin expression in Mp+/+ and Mp–/– embryoid bodies analyzed by immunostaining. Arrowhead shows the lumen in the Mp+/+ EB; arrow shows the endodermal cells which express maspin. Note no maspin protein is detected in Mp–/– EB. (C) GATA4 immunostaining of Mp+/+ and Mp–/– EBs. Arrows indicate the endodermal cells and arrowhead indicates the lumen of the Mp+/+ EBs. (D) HNF4 expression (fast-red labeled cells) of Mp+/+ and Mp–/– EBs analyzed by in situ hybridization. (E) Oct4 immunostaining of Mp+/+ and Mp–/– EBs.