Figure 2.

Transient overexpression of Dax1 does not drive differentiation. A, mES cells in 10-cm plates were transfected with 10 μg pEGFP Dax1 and 48 h later sorted for GFP-positive and -negative cells. Cells were harvested for QPCR analysis. As positive controls, cells differentiated by retinoic acid (RA) treatment or withdrawal of LIF (−LIF) were also subjected to QPCR analysis (as described in Materials and Methods; RA: Cdx2, Pax6, Rhox6, Foxa2, Fgfr2; -LIF: T, Fgf5). B, Sorted cells were replated and AP staining was performed. Percent of AP-positive cells was determined by Metamorph software.