(A) Allele-specific RT-PCR analysis. All RT- reactions were negative (not shown). Xist d0 samples (asterisks) were 10-fold overloaded to visualize low expression.
(B) Realtime RT-PCR of indicated transcripts, each normalized to β-actin.
(C) Immuno-RNA FISH for Xist and H3-3meK27 domains (arrowheads) on d10.
(D) Frequency of aberrant H3-3meK27 enrichment in the Xist+ subpopulation of indicated cell lines. n=100-150.
(E) Model: intersection of RNAi and XCI.
(F) Methylation-sensitive restriction analysis of the Xist promoter. Genomic DNA was digested with EcoRV or EcoRV+HpaII. % uncut (methylated) DNA at HpaII is plotted.