EMSA of mouse PXR proximal promoter.

<p>An end-labeled 134 bp promoter fragment (−297/−163) was incubated with AML-12 whole cell lysate. Lane <b>1</b>: free probe; Lanes <b>2, 3</b> & <b>4</b>: binding reactions performed with increasing amount of 3 μg, 6 μg and 12 μg of AML-12 lysate respectively. DNA-protein complexes are shown with arrows.</p

In silico analysis of mouse PXR proximal promoter.

<p>The nucleotide sequence of the 5′ flanking genomic DNA of the mouse PXR gene is shown along with consensus sequences for potential DNA-protein binding sites. Transcription initiation site, +1, is denoted by an arrow. PU.1, purine rich box binding element; Ets-1, Ets binding element; GR, glucocorticoid receptor binding element; ER, estrogen receptor binding element; VDR, vitamin D receptor binding element; COUP-TF, chicken ovalbumin upstream promoter binding element; NF-AT, nuclear factor of activated T cells binding element; AP-1, activator protein binding element; HNF1/3/4, hepatocyte nuclear factor binding element; AR, androgen receptor binding element; RXR/RAR, retinoid X/acid receptor binding site; T3R, thyroid hormone receptor binding element; PPAR, peroxisome proliferator-activated receptor binding element; GATA, gata binding element; LEF, lymphoid enhancer factor binding element; LyF, lymphoid factor binding element; c-Myb, myb (myeloblast) binding element; NF-1, nuclear factor binding element. Region marked with red color shows putative binding sites for different transcription factors and this region is further used for EMSA analysis.</p

A typical ChIP assay showing binding of Ets-1 and LEF/β-catenin to mouse PXR proximal promoter.

<p>Lane<b> 1</b> denotes PCR amplification of input DNA; lane<b> 2</b> shows PCR amplification of DNA immunoprecipitated using pre-immune serum (control IgG) and lanes<b> 3 & 6</b> represent PCR amplification of DNA immunoprecipitated by Ets-1 and β-catenin antibodies. No DNA was immunoprecipitated using PU.1 and Sp-1 antibodies (lanes<b> 4 & 5</b>).</p