Research Article

Alteration of Differentiation Potentials by Modulating GATA Transcription Factors in Murine Embryonic Stem Cells

Figure 1

Endoderm lineage differentiation of ES cells in vitro. Approximately 1 × 106 ES cells of wildtype or deficient in one of GATA factor were seeded on 100 mm plates as a monolayer culture (m) or cultured in suspension to allow cell aggregation to form spheroids (s). The cells were also treated with 1 μM retinoic acid (RA) or DMSO control. Following a 4-day culture period, cell lysates from monolayer (a) or from spheroids (b) were prepared for Western blotting analysis. (c) mRNA was prepared for Northern blot analysis. (d) Preservation of Oct-3/4 protein in embryoid bodies: embryonic stem cells were cultured in medium lacking LIF in suspension to allow the formation of cell aggregates. The embryoid bodies from a 4-day suspension culture were fixed in formalin, embedded in paraffin, sectioned, and immunostained for Oct-3/4 protein. Representative stainings of two embryoid bodies are shown (left panel, 40×, and right panel, 200×).
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