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-treatment of cells with vitamin D and ligand presence on factor binding were carried out prior to performing in vitro footprint experiments or electrophoretic mobility shift assays (EMSAs) involving competitors and antibodies. For each sample, nonspecific

-palindromic ERE from the human pS2 gene in electrophoretic mobility shift assay (EMSA) experiments ( Wood et al. 1998 ). Limited protease digestion of ERα bound to vit A2 ERE or the non-palindromic pS2, vitellogenin B1, and oxytocin EREs revealed different sized

°C with intensifying screens and the positions of the footprints determined. Electrophoretic mobility shift assay (EMSA) Synthetic sense and antisense oligonucleotides (Table 1 ) were used as probes in gel-shift and

concentrations were determined with the BCA protein assay system (Pierce Chemical Co., Montluçon, France). Electrophoretic mobility shift assay (EMSA) Nuclear extracts from GT1–7 and HEK293 cells were prepared as previously

yielding nuclear extracts for further processing. Protein concentrations of nuclear extracts were measured with the Roti-Nanoquant protein assay (Roth, Karlsruhe, Germany). Electrophoretic mobility shift assay (EMSA

aliquoted and stored at −135 °C until used. Protein concentration was measured with the bicinchoninic acid (BCA) protein assay kit from Pierce. Electrophoresis mobililty shift assay (EMSA) Double-stranded oligonucleotides

Results section and the figure legends, cells were processed for electrophoretic mobility shift assay (EMSA) or western blot analysis of phospho ERK1/2. Electrophoretic mobility shift assay Nuclear extracts from H32

shift assay (EMSA) Nuclear extracts from HEK293 cells with or without Dex treatment were prepared and EMSA was carried out as previously described ( Zhao et al . 1999 ). Briefly, oligonucleotides corresponding to the putative glucocorticoid response

for 30 s, 58 °C for 30 s and 72 °C for 30 s for a total of 35 cycles. In vitro transcription and translation, and electrophoretic mobility shift assay (EMSA) PGC-1α and ERRγ were transcribed and translated in vitro

mutation of the AP1 site located 52 bp downstream from the ERE decreases the estrogen response of the gene ( Barkhem et al. 2002 ). Multiple copies of EREs also influence the estrogen response of a target gene. Electrophoretic mobility shift assay (EMSA