Fig. 2
From: Ino80 is essential for proximal-distal axis asymmetry in part by regulating Bmp4 expression
Ino80 KO ESCs are viable, but differentiate when maintained in a metastable pluripotent state. a Microscopic analysis of wild-type and Ino80 KO pre-implantation E3.5 blastocysts. Blastocysts were allowed to outgrow for seven days onto gelatinized plates in media containing serum + LIF. Wild-type and Ino80 KO are at identical magnification. Red arrows designate ESC colony outgrowth. b Western analysis of Ino80 protein expression in wild-type and Ino80 KO ESCs using a custom Ino80 antibody. Ponceau S was used as a loading control. c Microscope analysis of wild-type and Ino80 KO ESCs grown in media containing serum + 2i + LIF or serum + LIF. Wild-type and Ino80 KO are at identical magnification. d Alkaline phosphatase (AP) activity in wild-type and Ino80 KO ESCs after plating at clonal density and maintained in media containing serum + 2i + LIF or serum + LIF. Wild-type and Ino80 KO are at identical magnification. e Percent AP positive colonies quantified from panel d and after Ino80 KO ESCs passaged in serum + LIF were recovered in serum + 2i + LIF. At least 50 colonies were quantified per group. f Quantitative RT-PCR analysis of gene expression in wild-type and Ino80 KO ESCs maintained in serum + 2i + LIF. (N = 3 biological replicates) g Quantitative RT-PCR analysis of gene expression in wild-type and Ino80 KO ESCs maintained in serum + LIF. (N = 3 biological replicates; * = t test p ≤ 0.05). KO knockout, ESC embryonic stem cells, E embryonic day, LIF leukemia inhibitory factor,