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. Author manuscript; available in PMC: 2013 Dec 20.
Published in final edited form as: Cell. 2013 Jun 20;153(7):1537–1551. doi: 10.1016/j.cell.2013.05.028

Figure 3. CTCF Overexpression Results in Female-Specific EB Outgrowth Defect.

Figure 3

(A) Dox-inducible bidirectional expression of CTCF-3xFLAG and GFP in rtTA-expressing MEF. Normal cell morphology shown by phase-contrast before (Dox—) and after (Dox+) induction of GFP and CTCF. FLAG and CTCF immunofluorescence shown. (B) Western blot of representative transgenic clones shows dox-induction of CTCF-3xFLAG in d0 ES cells (left panel) and d1 female ES cells (right panel). (C) CTCF overexpression results in female-specific defect in outgrowth up to d13. Bright field images show EB outgrowth. GFP indicates transgene (CTCF) overexpression. Note GFP-positive cells in male, but not female, outgrowth in Dox+ condition. After induction, GFP signals are confined to the EB center. (D) Female control, ctrl#5, carries a silenced transgene and grows normally, whereas overexpressers, clones #2 and #1, fail to outgrow. d5 images shown.