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Islet binding was verified by immunostaining the same sections to detect insulin.
In parallel studies, the specific cell uptake due to σ receptor binding was verified by the presence of a variety of σ ligands including -pentazocine (σ1), haloperidol (non-selective σ1/σ2) and unlabelled SIG343 and SIG353 (σ2).
Concentration-dependent binding was verified by a linear response when plotting on double-logarithmic scale.
For each of the peptides HLA-A*0201 HLA-A*0201s verified bindingmpetition assay.
The presence of anti-CD15-SPIONs binding was verified by Prussian blue staining.
HuR binding was verified by HuR immunoprecipitations, and analyzed by quantitative RT-PCR and biotin pull-downs.
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The identity of the protein and the specificity of the binding were verified by competition with a point mutated mb-1/EBF site and by the inclusion of an EBF specific antibody into the reaction mixture.
Aptamer binding was verified 72 h postsilencing.
Subsequently, the binding of FOXF2 on the TWIST1 promoter region containing or lacking the putative binding site was verified by ChIP assay in MCF-10A cells.
N-6 lacks such a carboxylate moiety, and therefore it should not require Arg316 for binding, which was verified by our mammalian one-hybrid assay showing that the mutation of Arg316 did not affect the inhibition of RXRα transactivation by N-6 (Fig. 4A).
The nucleotide sequence of each binding site was verified by crosschecking with the UCSC database, and the specific genomic coordinates for each sequence were recorded.
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