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. 2015 Jul 29;290(37):22460–22473. doi: 10.1074/jbc.M115.675595

FIGURE 3.

FIGURE 3.

Etv4/5 dKO ES cells maintain the undifferentiated state in N2B27 serum-free medium plus two inhibitors. A, morphological signatures of PE9 (WT) and PE15-2 (Etv4/5 dKO) ES cells. PE9 and PE15-2 ES cells were cultured in either a regular serum- and LIF-containing medium or a chemically defined medium (DMEM/F-12 with N2B27 supplement plus 2i (mitogen-activated protein kinase/extracellular signal-regulated kinase (PD0325901) and glycogen synthase kinase 3 (CHIR99021) inhibitors) (2i condition)). Morphological signatures were observed. B, expression of Etv4 and Etv5 in the 2i condition. Expression of Etv4 and Etv5 in the conditions of A in PE9 and PE15-2 ES cells was examined by quantitative RT-PCR. C, expression of self-renewal marker genes in PE9 and PE15-2 ES cells. Expression of the indicated self-renewal marker genes in the conditions of A in PE9 and PE15-2 ES cells was examined by quantitative RT-PCR. All samples were analyzed in triplicate, and the data were normalized to Gapdh expression. Error bars, S.D.