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. 2013 Dec 31;8(12):e83426. doi: 10.1371/journal.pone.0083426

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© 2013 Hoffmann et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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EMSA bands were obtained incubating either the Cy5 labeled probes IVS1+1505G (A, lane 1) or SP1/SP3 consensus (B, lane 1) with nuclear extracts from HIB1b cells followed by native PAGE. Non-labeled competitors IVS1+1505G, IVS1+1504A and SP1/3 consensus were added to the binding reaction along with labeled probe where indicated. Different spacing between the complexes and a non-SP complex formed with the IVS1+1505G probe (arrows in (A), competition with SP1/3 consensus) hint to different complex compositions. (C) Supershift experiments by addition of antibodies against SP1, SP3, PPARγ and RXRα to test the identity of the proteins binding to the IVS1+1505G element. A representative experiment of 3 independent repetitions is shown in C.