Figure 2
From: The Mbd1-Atf7ip-Setdb1 pathway contributes to the maintenance of X chromosome inactivation
Nuclear localization of ATF7IP relative to Xist RNA, H3K27me3, and H3K9me2 during the initiation and maintenance phases of XCI. (A) Undifferentiated male mouse ESCs carrying a tet-inducible promoter at the endogenous Xist locus were analyzed by FISH for Xist RNA (red) and immunostaining for H3K9me2 and ATF7IP, respectively (green), 21 h after induction of Xist expression with doxycycline. DAPI was used to detect nuclei. Representative images are shown and the numbers on the right of the merged images indicates the percentage of cells (n = 100 per sample) with H3K9me2 or ATF7IP enrichment within the Xist RNA-demarcated X chromosome. (B) Similar to (A), except that female mouse ESCs after 5 days of retinoic acid (RA) differentiation were analyzed by H3K27me3/H3K9me2 co-immunostaining or Xist RNA FISH in combination with ATF7IP immunostaining. (C) Similar to (A), except that female MEFs were stained for H3K9me2, H3K27me3, ATF7IP, and Xist RNA in the indicated combinations.