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Fig. 3. The Xist gene on X^pA is dysfunctional. (A) The number of male and female pups born to wild-type females crossed with males hemizygous for the Tsix^pA allele. The sex ratio was extremely biased toward males. One of two females turned out to be XO. (B) The number of male and female embryos recovered at E7.5 from wild-type females crossed with X^pAY males. Two of the embryos were genotyped as XX^pAY. (C) Gross morphology of the typical XX^pA embryo recovered at E7.5 is shown together with that of a male littermate. This phenotype is quite similar to that of females that inherit an Xist-deficient X from the father. e, embryonic ectoderm; ec ectoplacental cone; rm, Reichert's membrane. Scale bar: 0.5 mm. (D) The numbers of male and female pups born to females heterozygous for Tsix^pA crossed with wild-type males are shown. The Tsix^pA allele is transmitted to both male and female pups at the expected ratio. (E) Females heterozygous for Tsix^pA were crossed with X^GFPY males and embryos recovered at E8.5 were examined for GFP expression. Although GFP fluorescence is uniformly observed in wild-type XX^GFP embryos because cells that did not select X^GFP as the inactive X are distributed throughout the body, X^pAX^GFP embryos are substantially negative for GFP, indicating that X^GFP is invariably inactivated. This demonstrates that the Xist gene on X^pA is dysfunctional. ys, yolk sac; am, amnion; al, allantois. Scale bar: 0.8 mm.

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