FIGURE 5.

Elf3 bound the Ets element in the AQP2 promoter in the mpkCCD cells. (A) Electrophoretic mobility shift assay. Nuclear extract from either V5-tagged Elf3 isoform 1 (left panel) or V5-tagged Elf3 isoform 2 (right panel) expressing mpkCCD cells was incubated with a ³²P isotope-labeled AQP2 promoter DNA (hot probe) and subjected to electrophoresis followed by autoradiography. A non-isotope labeled competitor AQP2 promoter DNA (cold probe) or a V5 antibody (at 1, 2, or 4 μg) was used to inhibit the interaction. A non-specific IgG was used as a control. (B) ChIP-PCR assay. Chromatin immunoprecipitation was done with a control (IgG), V5 tag, or Elf3 antibody before PCR analysis with primer sets framing the indicated regions in the AQP2 promoter. N is a no template negative PCR control.