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

Figure 3

Primitive endoderm differentiation and the absence of the basement membrane in GATA5-deficient embryoid bodies. Wildtype and GATA5 (−/−) ES cells were cultured in medium lacking LIF in suspension to allow the formation of cell aggregates on Petri dishes, with or without retinoic acid (1 $\mu$M) for 4 days. The embryoid bodies from a 4-day suspension culture were fixed, embedded in paraffin, sectioned, and subjected to PAS staining to detect the basement membrane. Representative stainings are shown (200$\times$). Spheroids of GATA4 (−/−) and GATA6 (−/−) ES cells were previously published [25] and showed that endoderm formation is impaired in GATA4 (−/−) ES cell spheroids without RA. Endoderm formation in GATA4 (−/−) ES cell spheroids can be restored by treatment with RA for 4 days. Spheroid of GATA6 (−/−) ES cells lacked endoderm formation that cannot be restored with RA treatment [25].