Figure 5. RNA FISH analysis of X-inactivation in Eed^m-/- and Eed^mz-/- blastocysts.

(A,B) RNA FISH detection of Xist RNA (green) and Rnf12 RNA (red) in representative Eed^m-/- and Eed^mz-/- female (A) and Eed^mz-/- male (B) blastocysts. Nuclei are stained blue with DAPI. Scale bars, 20 µm. Individual nuclei of representative categories of stain are shown to the right of each embryo. Eed^fl/fl female data from Figure 3D shown for comparison. Mutant female embryos ranged in size from 46 to 80 nuclei. Fully developed mutant male embryos ranged in size from 53 to 110 nuclei. Delayed mutant male embryos ranged in size from 30 to 40 nuclei. Bar plot shows percentage of nuclei in each embryo with Xist RNA coats and/or Rnf12 RNA expression. Each bar represents an individual embryo and embryo numbers under the bars correspond to the same female embryos plotted in Figure 4A. *, p<0.05; **, p<0.01, Two-tailed Student’s T-test, between Eed^m-/- and Eed^fl/fl, or Eed^mz-/- and Eed^fl/fl. (C) Data showing the number of Eed^m-/- embryos which can live to term compared to Eed^fl/fl embryos. WT, wild-type. Table shows Eed^m-/- litters sired by Mus musculus-derived male or Mus molossinus-derived male. Male Eed^m-/- offspring are underrepresented compared to females, p=0.02, Two-tailed Student’s T-test.