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. 2016 Mar 7;113(12):3197–3202. doi: 10.1073/pnas.1523538113

Fig. S8.

Fig. S8.

Rnf12 overexpression does not induce an ectopic iXCI in male embryos. (A) Relative expression levels of Rnf12 and Xist in male embryos overexpressing Rnf12 (Rnf12-OX) and Egfp (Egfp-CTL) during the preimplantation stage. Each diamond represents the expression value detected by single embryo qRT-PCR. Horizontal gray lines represent mean values. The number of embryos in each group is indicated. (B) RNA-FISH analyses of Xist noncoding RNA in Rnf12-OX and Egfp-CTL male embryos. (C) H3K27me3 immunostaining in Rnf12-OX and Egfp-CTL male embryos; trophoblast cells were labeled with CDX2. (D) Relative mRNA expression levels of pluripotency factors (Nanog, Oct4, and Sox2), Rnf12, and Xist in pooled IVO and IVF female embryos. Sox2 was not detected at the eight-cell stage. (E) NANOG immunostaining in IVO and IVF female blastocysts. White arrows indicate the mislocalization of NANOG in trophoblast cells. (Scale bar, 50 μm.) *P < 0.05.