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. Author manuscript; available in PMC: 2015 Dec 18.
Published in final edited form as: Cell. 2014 Dec 18;159(7):1681–1697. doi: 10.1016/j.cell.2014.11.040

Figure 3. Xi Features in Relation to the Sequential Expression of Pluripotency Factors during Reprogramming.

Figure 3

(A) Quantitation of an immunostaining analysis of NANOG, XiEZH2+, and REX1, presenting the proportion of NANOG+ colonies with XiEZH2+ and/or REX1 expression at indicated time points. Right, representative immunostaining images for EZH2 (magenta in the merge), NANOG (red), and REX1 (green).

(B) As (A) for NANOG, XiEZH2+, and DPPA4.

(C) Quantitation of an immunoFISH analysis for PECAM1 and Xist RNA displaying the proportion of PECAM1+ cells with XiXist+.

(D) Representative immunostaining image for EZH2 (magenta in the merge), NANOG (red), and PECAM1 (green), demonstrating the absence of XiEZH2+ in PE-CAM1+ cells.

(E) (i) Quantitation of mono- and biallelic expression of the X-linked gene Atrx within cells of small (<12 cells) and large (>20 cells) DPPA4+ colonies and representative images from the DPPA4 (magenta) and Atrx (green) immunoFISH staining. Arrowheads indicate Atrx nascent transcription signals. (ii) Quantification of biallelic Atrx expression in PECAM1+/− cells.

(F) Summary of reprogramming stages identified in this figure as in Figure 2E.

See also Figure S3.