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. 2006 Jun 26;103(27):10294–10299. doi: 10.1073/pnas.0506945103

Fig. 4.

Fig. 4.

BMP signaling promotes the mesoderm specification of ES cells. (A) Flow-cytometric analysis of T-positive cells in T-EGFP ES cells cultured for three passages with 400 units/ml LIF under conditions of inhibition (noggin) or activation (BMP2, BMP4, and BMP7) of BMP signaling. Bar shows mean ± SD (n = 4). (B) Flow-cytometric analysis of T-positive cells produced from purified T-positive cells cultured with 400 units/ml LIF under conditions of inhibition or activation of BMP signaling. Inhibition of endogenous BMP signaling by noggin decreased the percentage of T-positive cells at a similar level of Nanog overexpression, whereas BMP activation increased the percentage of T-positive cells. Bar shows mean ± SD (n = 4). (C) RT-PCR analyses in ES cells cultured with 400 units/ml LIF with or without addition of BMP7 show that BMP-dependent Id1 expression is negatively regulated by overexpressing Nanog and enhanced when Nanog function is down-regulated. (D) A reporter construct of −1147Id1-Luc containing the Smad-binding sites, but not −927Id1-Luc, was activated in a BMP-dependent manner in ES cells cultured with 400 units/ml LIF. Nanog and inhibitory Smads (Smad6 and Smad7) down-regulated −1147Id1-Luc activity in a similar manner. Bars show mean ± SD (n = 4).