Search Results

. Cell fractionation and electrophoretic mobility shift assays To test for factors binding to the putative transcription factor-binding site, electrophoretic mobility shift assays (EMSAs) were carried out essentially as described previously ( Taylor et

phenylmethylsulfonylfluorid, 1 mM benzamidine, 30 mg/ml leupeptin, 5 mg/ml aprotinin, 5 mg/ml pepstatin). Nuclear extract was aliquoted and stored at −80 °C. Electrophoretic mobility shift assays (EMSAs) were performed in a 20 μl binding reaction containing 10 μg of the

-stranded oligonucleotides (Sangon, Shanghai, China) were designed and synthesized. The DNA binding activity of C/EBPβ protein was detected by LightShift Chemiluminescent EMSA Kit (Thermo Fisher Scientific, Waltham, MA, USA). Ten microgram pig abdominal fat nuclear extract

transfection experiments to test for functional activity. Electrophoretic mobility shift assay (EMSA) was also used to study protein–DNA interactions at the GC-responsive region of the hDUSP1 promoter region. Materials and methods Cells and growth conditions

assayed. The promoterless pGL3-Basic vector was included as a control in the transfection experiments and the results of the luciferase activity were calculated relative to the activity of pGL3-Basic. Electrophoretic mobility shift assay (EMSA) EMSA was

system using the ImageQuant software version 1.1 (Molecular Dynamics, Amersham Biosciences). Granulosa cell nuclear extracts and electrophoretic mobility shift assays (EMSAs) Equine granulosa cells were obtained from

1130 ATACTCAGAATAGATTATG siERα_3 FRD CAUCUUGCUUAAUUCUGGAdTdT 1508 CATCTTGCTTAATTCTGGA siERα_3 REV UCCAGAAUUAAGCAAGAUGdTdT 1508 TCCAGAATTAAGCAAGATG Electrophoretic mobility shift assay (EMSA) Cells were

luciferase reporter assays could follow a previously described method ( Zhang et al. 2018 ) . Electrophoretic mobility shift assays For electrophoretic mobility shift assays (EMSAs), nuclear proteins (NPs) were extracted from bovine longissimus

(EMSA) BgRXR cDNA was cloned into XhoI/KpnI digested pTL1 (a modified version of pSG5; Stratagene, La Jolla, CA, USA), for transient transfection assays and in vitro translation. Vectors expressing potential heterodimer partners were as

. Products were analyzed by 1.5% agarose gel electrophoresis and quantified by real-time PCR. Fold enrichment = 2 Ct (IgG)−Ct (HIF1α) . Electrophoretic mobility shift assay (EMSA) and supershift assays EMSA was carried out to determine HIF1