FIG. 3.

Proliferation rates and BMP signaling are unaltered in Ofd1^Gt EBs. (A) Phospho-histone H3 staining in wild-type and Ofd1^Gt EB sections differentiated for 7 days. Scale bar 20 μm. (B) Quantification of phospho-histone H3 stained cells per given area of wild-type and Ofd1^Gt EBs after 4, 7, and 10 days of differentiation. (C) BrdU incorporation as measured by colorimetric absorbance confirming similar proliferation in wild-type and Ofd1^Gt EBs grown for 7 days. (D) Immunoblot of wild-type and Ofd1^Gt cells during differentiation probed for phospho-Smad 1/5 and β-actin. (E) Neural specification as measured by Sox1 expression levels in wild-type and Ofd1^Gt EBs treated with BMP4 or Noggin, a BMP antagonist. (F) Assessment of Smad1 knockdown in wild-type and Ofd1^Gt EBs containing Smad1 RNAi lentivirus. (G) Expression of Sox1 and Nestin neural markers in wild-type and Ofd1^Gt EBs treated with Smad1 RNAi. Asterisk indicates statistical significance of p ≤ 0.05. BMP, bone morphogenic protein; BrdU, bromodeoxyuridine. Color images available online at www.liebertonline.com/scd