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. Author manuscript; available in PMC: 2007 Oct 9.
Published in final edited form as: FEBS Lett. 2007 May 11;581(14):2633–2640. doi: 10.1016/j.febslet.2007.05.006

Figure 5.

Figure 5

Zfp496 can bind DNA and activate SAAB reporter gene constructs. A. Representative GMSA using GST-Zfp496 and the DNA-binding motif #18 selected by SAAB. Lane 1: #18 probe alone. Lane 2: Probe plus 2.5 μg GST only. Lanes 3 and 4: Probe plus 0.5 μg and 1.5 μg GST-Zfp496, respectively. The binding mixture was subjected to non-denatured PAGE and autoradiographed. B. Transient transfection assays were performed in 10T1/2 cells cultured in 6-well plates using the reporter (50ng) and Zfp496 (50ng, 100ng, 200ng) using Lipofectamine 2000. Relative luciferase activity was calculated as in Fig. 3. C. The oligonucleotide sequences of the high affinity DNA-binding sites selected by SAAB are shown. The underlined letters indicate binding sites of homeobox proteins. * indicates the oligonucleotides that were identified twice each. D. Transfection assays were performed in 10T1/2 cells in 12-well plates using the reporter (300ng), Zfp496 (400ng) and JMJ (250ng, 500ng, 1000ng) using Lipofectamine 2000. Each graph in B and C represents at least three independent experiments of duplicate plates.