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. Author manuscript; available in PMC: 2009 Sep 4.
Published in final edited form as: Dev Dyn. 2006 May;235(5):1358–1370. doi: 10.1002/dvdy.20722

Fig. 4. Stat1 binds to the Mtf6 DCE region of the Tcfap2a FNP/LBM enhancer.

Fig. 4

(A) Schematic representation of the Mtf6 DCE subfragment of the FNP/LBM enhancer. The sequence of Probe 45, a highly conserved sub-fragment of the Mtf6 DCE region, contains several putative binding sites for known classes of transcriptional regulators including; bHLH, Ets, STAT, and homeodomain proteins. Consensus sequences for these putative binding factors are shown. SIE is a naturally occurring binding site for Stat1. (B) EMSA experiment using radiolabelled Probe 45 either with (+) or without (-) protein extract from Stage 25 chick FNP. The two specific complexes formed are indicated (arrows). Lanes also contained either no specific cold competitor DNA (-) or various cold competitors containing particular consensus sequences (listed in A and indicated above the relevant lanes) at 10-fold and 100-fold excess in adjacent lanes. Only the cold competitor containing the STAT consensus competed effectively for the upper shift. (C) Stat1 in extracts from U-937 cells treated with IFNa (oval) and E10.5 mouse head extracts (rectangle) binds to both SIE and Probe 45 (lower arrows). The Stat1 shifts on both probes and with both types of extracts are super-shifted in the presence of a Stat1-specific antibody (upper arrows).