Fig. 6. Histone modifications and Xist accumulation in female XEN and TS cells. (A,B) Representative images show immunofluorescent detection of Eed, Ezh2 and histone modifications H3 di/tri-methyl K27 and H3 acetyl K9, combined with Xist RNA FISH. Immunodetections were performed using Alexa 568-conjugated secondary antibodies (red). Xist RNA was detected using a Spectrum Green-labeled FISH probe (green). XEN cells (A) show no Eed and Ezh2 enrichment, and weak H3 di/tri-meK27 enrichment on the Xist-coated X chromosome, but they do show depletion for H3 acK9. By contrast, undifferentiated TS cells (B) show enrichment for Eed, Ezh2 and H3 di/tri-meK27 on the Xist-coated X chromosome. Depletion for H3 acK9 was observed, as in XEN cells.