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. 2009 Jan 21;106(4):1122–1127. doi: 10.1073/pnas.0812210106

Fig. 4.

Fig. 4.

X-inactivation in female hybrid CAST/Ei xC57BL/6 Dicer KO ES cells. (A) PCR analysis of genomic DNA for the presence of dcrfl, dcrΔ and CAST and C57BL/6 SNP alleles (SNP RS29080486). (B) qRT-PCR for Oct4 and Nanog mRNA before (d0) and after RA exposure (d2 and d6). (C) DNA FISH indicating the presence of 2 X chromosomes in differentiating ES cells 8 days after RA exposure using X paint (green) and XIC probe (red). (D) Left images, FISH for Xist RNA (green) with a single-stranded RNA probe in RA-treated dcrfl/fl and dcrΔ/Δ cultures at day 6 of differentiation. Right images depict merged images of Xist RNA (green) and nuclear Hoechst staining (blue). (E) Strand-specific qRT-PCR for Xist RNA before (d0) and after RA exposure (d2 and d4). (F) Combined IF-FISH for Xist RNA (green, Left images) and Ezh2 (red, Middle images) in RA-treated dcrfl/fl and dcrΔ/Δ cultures at day 2 of differentiation. Right images depict merged images of Xist RNA (green), Ezh2 (red) and nuclear Hoechst staining (blue). Arrows indicate regions of Xist/Ezh2 co-localization.