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. Author manuscript; available in PMC: 2011 Nov 15.
Published in final edited form as: Cancer Res. 2010 Nov 2;70(22):9197–9206. doi: 10.1158/0008-5472.CAN-10-1568

Figure 6.

Figure 6

DLX5 binds to and transactivates the IRS-2 promoter. (A) Luciferase assay was performed by co-transfecting pGL3-IRS-2 promoter with GFP or with pMSCV expressing FLAG tagged full-length (FL) DLX5 or DLX5 with truncated C terminus (ΔC’), N terminus (ΔN’) or terminal Homeobox domain-C (ΔH-C). (B) Western blot analysis demonstrates the expression of FLAG tagged full-length and truncated DLX5 by using a anti-FLAG antibody. (C) Gel shift assay was carried out to analyze the interaction between recombinant DLX5 protein and its binding consensus in the IRS-2 promoter. (D) DLX5 immunoprecipitated from nuclei of IGR-OV1 cells (IGR) also binds to IRS-2 promoter. IP from SKOV3 cells (SK) was used as the negative control. (E) Luciferase assay was performed by co-transfecting the mutant pGL3-IRS-2 promoter with DLX5. (F) Oligos with mutant DLX5 binding sequence was used for the gel shift assay. (G) Chromatin immunoprecipitation assay performed to determine the interaction of DLX5 and IRS-2 promoter at the endogenous level.