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The ChIP assays revealed that endogenous and overexpressed Runx2 binds to mTOR promoter and that this binding is reduced by Runx2 knockdown.
Given that SP1 and SP3 compete for binding sites, it is possible that reduced binding of endogenous SP1 to the methylated C and T alleles combined with the increased SP3/SP1 ratio may increase the number of cells in which SP3 can bind over rs143383, even though the affinity of SP3 binding is reduced by methylation.
In kidneys of Agtr1A mice, AT1-specific binding is virtually undetectable and renal AT1-specific binding is reduced by approximately 50% in Agtr1A heterozygotes.
The reduced free energy between MB target duplexes and unbound MB, creates an environment where the energetic choice of target-specific binding is reduced by a single base mismatch [16].
In cells over-expressing dis1p, ase1p binding is reduced by the dynamics of dis1p leading to ase1p-GFP being more dynamic and dispersed in the IMAs.
Thus, in the sequential titration of MsbA with ATP or its non-hydrolysable analogue p[NH]ppA, and lipid A, it was been observed that the affinity of lipid A binding is reduced by only 3- to 8-fold when nucleotide is pre-bound, which is unlikely to be large enough to trigger dissociation of the lipid.
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In their study, sequential mutational analysis revealed that FOXL2 binding was reduced by mutation of any of the 12 consecutive base pairs at their site.
Non-specific binding was reduced by incubating the membrane in a blocking buffer (20 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.1% Tween 20, 1 µM protease inhibitors, 5 mM NaF, and 1 mM Na3VO4) containing 10% non-fat dried milk for 1 hour.
Nonspecific binding was reduced by incubation in 1% BSA for 60 min.
Unspecific binding was reduced by incubation in 10% Roti®-Immunoblock (Roth, Karlsruhe, Germany).
When the cells had been treated with compactin to deplete cholesterol, IPFO* binding was reduced by 92%.
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binding is counterbalanced by
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