FIGURE 2.

RNA interference knockdown of mSin3A-HDAC complex members affects Nanog expression without inducing differentiation. A, Western blot showing levels of mSin3A, HDAC1, and HDAC2 proteins in siRNA-treated ES cells. Lamin siRNA was used as a negative control. Glyceraldehyde-3-phosphate dehydrogenase was used as a loading control. B, levels of Nanog protein (top) and mRNA (bottom) in response to knockdown of mSin3A and associated HDAC members; 96 h after indicated siRNA treatment. Relative protein levels were quantified by image densitometry. Values for Nanog mRNA and protein were normalized to Lamin siRNA. C, mRNA expression levels of pluripotency-related genes (Oct4, Nanog, Sox2, and Rex1), mesodermal lineage markers (Bmp4 and Hand1), ectodermal lineage marker (Nestin), endodermal lineage markers (Gata4 and Sox17), trophectodermal lineage markers (Cdx2 and Eomes), and primitive ectoderm lineage (Fgf5) after siRNA treatment for Lamin (control) or mSin3A.