Fig. 3

Ino80 is essential for cell and tissue differentiation. a Low-magnification analysis of wild-type and Ino80 KO ESC differentiation as teratomas in NOD/SCID mice. Scale bar = 500 μm. b High-magnification analysis of teratomas derived from wild-type ESCs showed evidence of (a’) blood islands, (b’) striated muscle, (c’) ciliated endoderm, (d’) neural rosettes, (e’) keratin pearls, and (f’) smooth muscle fields when sectioned and stained with hematoxylin and eosin (H + E). Teratomas formed from Ino80 KO ESCs are composed of (g’) undifferentiated tissue surrounded by a (h’) thin epithelial cell layer. Scale bar = 75 μm. c Analysis of 9-day embryoid bodies derived from wild-type and Ino80 KO ESCs. Frozen sections of embryoid bodies derived from wild-type ESCs stained with H + E showed evidence of dark staining endoderm (Endoderm) and evidence of internal cavitation (Cav) as previously described [24]. Embryoid bodies from Ino80 KO ESCs show evidence of disorganized endoderm and other differentiated cell types. Scale bar = 200 μm. d Gene expression analysis of embryoid bodies derived from either wild-type or Ino80 KO ESCs differentiated over nine days. Quantitative RT-PCR was used to measure the expression of a variety of differentiation markers, including Gata6, Gata4, Foxa2, Hnf4a, Wnt3, Fgf8, T, Fgf5, Bmp4, and Nodal, and pluripotency markers, including Nanog, Klf4, Sox2 and Oct4. (Representative of N = 3 biological replicates; * = t test p ≤ 0.05). KO knockout, ESC embryonic stem cell