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. 2012 Feb 28;40(12):5718–5730. doi: 10.1093/nar/gks177

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© The Author(s) 2012. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — NHTF binding to the NHTF gene cluster and effect of DMP19 on its specificity. (A) The NHTF gene cluster encompasses a 5′-untranslated region along with the coding region. (B) The NHTF 5′-untranslated region was cut into four fragments with similar length (∼40-mer) and incubated with NHTF. Only fragment 3 was shifted after incubation with NHTF. (C) EMSA results showed that within fragment 3, only the 25-mer DNA fragment that included the un-mutated TGTNAN₁₁TNACA pseudo-dyad axis sequence produced the specific DNA shift. (D) Addition of the DNA mimic DMP19 reduces the specific DNA binding of NHTF in a dosage-dependent manner. The addition of the other two negatively charged proteins BSA and DMP12 had no effect. The DNA concentration used in all of the EMSA assays is 1 µM.