Figure 4

Differentiated EBs were analyzed by either immunocytochemistry or RT-PCR to the indicated molecular markers. (A) Immunostaining with goat anti-human SOX17 (Red), is contrasted with Fluoro Nissl nuclear staining (Green). (B) Immunostaining with goat anti-human GATA6 (Red), is contrasted with DAPI nuclear staining (Blue). (C) Immunostaining with goat anti-human brachyury (Red), is contrasted with DAPI nuclear staining (Blue). (D) Immunostaining with mouse anti-human GATA1 (Red). Note that each antibody recognizes subsets of EB cells. Scale bars = 100 μm. (E) The differentiation status of EB is detected by RT-PCR using different germ layer cell markers. Selected endoderm markers AFP, FoxA2; mesoderm markers Hand1, MSX1 and ectoderm marker Msl1 were all highly expressed in the EB samples while their expression was either undetectable or at low level in the ES samples. G3PDH was a positive control showing similar amount of RNA samples were used for analysis.