Fig. 1

Absence of H3K27me3 enrichment and Xist RNA coating on the inactive X chromosome in Eed ^–/– TSCs. a Top, diagram of conditional Eed mutation. Bottom left, gel image of genotyping PCR. Bottom right, RT-PCR detection of Eed RNA in Eed ^fl/–, Eed ^–/–, and Eed ^+/+ samples. The location of PCR and RT-PCR primers are depicted in the diagram above in blue and green, respectively. b Loss of Eed exon 7 and reduction of mRNA in Eed ^–/– line by RNA-seq. c Detection of EED (green) and H3K27me3 (red) by immunofluorescence (IF) and Xist RNA (white) by RNA FISH in Eed ^fl/fl and Eed ^–/– TSCs. Nuclei are stained blue with DAPI. Scale bar, 2 μm. Right, quantifications of numbers of nuclei with H3K27me3 enrichment and Xist RNA coating in Eed ^fl/fl and Eed ^–/– TSCs. d Absence of Xist RNA expression in Eed ^–/– TSCs by RNA-seq. e Enrichment of H3K27me3 at Xist promoter region on the inactive paternal-X in WT TSCs