Fig. 1.
Xist RNA coating and Polycomb protein recruitment in male Dicer KO ES cells forced to express Xist upon dox addition. (A) Upper images depict representative RNA FISH stainings for Xist (green) by using a strand-specific RNA probe in male dcrfl/fl and dcrΔ/Δ ES cells carrying the tet-inducible Xist allele, upon dox-induction of Xist. Lower images represent the merge with Hoechst staining of nuclei (blue). (B) Quantitation of the fraction of cells displaying robust Xist RNA coating. (C) IF/RNA FISH colocalization of Xist with Eed (D) Ezh2, and (E) trimethyl-H3K27, respectively, in dox-induced ES cells. Left images depict Xist RNA detected by FISH with a strand-specific RNA probe (green). Middle images depict immunostaining of indicated antigens (red). Right images depict merged images of RNA FISH, immunofluorescence and Hoechst staining. (F) Graphs depict quantitation of the percentage of cells with Xist RNA coating of the X chromosome that recruit the indicated PcG proteins and H3-m3K27 to the Xist RNA associated area.