Isolation of UTF1 homozygous mutant ESCs from blastocysts.

<p>(A) Western blot analyses of pluripotency marker proteins in ESCs generated by outgrowth of <i>UTF1</i> homozygous and heterozygous mutant blastocysts. (B) Comparison of the differentiation propensity between <i>UTF1</i> homozygous and heterozygous mutant ESCs. <i>UTF1</i> homozygous and heterozygous mutant ESCs were induced to differentiate by embryoid body formation. RNAs were prepared at the indicated days. cDNAs generated by reverse transcription were used to examine the levels of differentiation marker gene expression by real-time PCR. Data represent the mean with SD (n = 3). (C) Western blot analyses of Arf tumor suppressor protein in <i>UTF1</i> heterozygous and homozygous mutant ESCs. (D) Chimeric mouse analyses of <i>UTF1</i> homozygous mutant ESCs. Fluorescent Kusabira orange-labeled <i>UTF1</i> homozygous mutant ESCs generated from a blastocyst were injected into blastocysts. Embryos were allowed to develop in a recipient female mouse. Left and right panels are bright-field and fluorescence images of a 9.5 dpc embryo recovered from the recipient mouse, respectively.