FIG. 6.

Deregulated mRNA and protein expression is evident in Dnmt[3a^−/−,3b^−/−] P75 ES cells. (A) Dnmt[3a^−/−,3b^−/−] P75 ES cells and wild-type and 3b3 rescued ES cells cultured in 100 U of LIF/ml and stained for alkaline phosphatase (Alk. Phos.), laminin B1, and Oct4 by immunocytochemistry. Ab, antibody. (B) RT-PCR analysis of Oct4, Xist, and Gapdh in wild-type, Dnmt[3a^−/−,3b^−/−] P75, and Dnmt1^−/− EBs after induction to differentiate by LIF withdrawal. diff., differentiation. (C) RT-PCR analysis of trophoblast (Tpbp; Pl-1), endoderm (Hnf4a; albumin), mesoderm (brachyury; βh1 globin and α-globin), and housekeeping (Hprt and β-tubulin) genes in differentiating EBs generated from wild-type, Dnmt[3a^−/−,3b^−/−] P75, and Dnmt1^−/− ES cells.