Fig 1: Hypoxia–reoxygenation (H-R) challenge elicits a significant increase in the association and binding of NF-?B at the miR-210 proximal promoter. (A–C) miR-210 promoter pull-down assay determining the quantitative enrichment of the p65 NF-?B and p50 NF-?B subunits in the reverse crosslinked miR-210 promoter pull-down fragment. Representative Western blots (A) and quantitative densitometric analysis (B,C) showing the relative quantitative abundance of p65 NF-?B and p50 NF-?B subunits in the miR-210 promoter pull-down lysates. (D) Tandem ChIP-ELOHA analysis showing the relative enrichment of p65 NF-?B at the ?B response element in the miR-210 proximal promoter. Data from the Western blot-coupled densitometric analysis are expressed as mean fold change ± S.D. from three biological replicates belonging to each experimental group (n = 3). Data from the ChIP-ELOHA analysis are expressed as fold enrichment by first correcting the p65 NF-?B-associated miR-210 promoter fragment ELOHA absorbance values to the respective inputs, followed by normalization to fold change values. Data from the ChIP-ELOHA analysis are depicted as mean fold enrichment ± S.D. from three technical replicates for each of the four biological replicates belonging to each experimental group (n = 4). **** p = 0.0001. S.D.: standard deviation.
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