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We compared the activated and repressed enhancer clusters for differences in preferential binding of specific TFs.
The SNP probes of our arrays will be particularly valuable in evaluating whether a disease-associated or trait-associated SNP alters the binding of specific TFs.
Because modification of histone tails in enhancer regions influences DNA accessibility, we wanted to determine if the binary regulation (activation or repression) of enhancers corresponds to the binding of specific TFs during EMT.
Binding of specific TFs was tested by competition EMSA and by antibody-supershift EMSA.
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Our results suggest that morphological divergence is not driven by subtle drift in specificity of TFs, but primarily caused by cis-regulatory changes (Carroll, 2000; Levine and Tjian, 2003), with some contribution from relatively large shifts in binding specificities of specific TFs.
Thus, not all detected putative gains of TFBSs in BY4741 or losses in other strains or species necessarily reflect real changes of TF binding, but might be the consequence of changes in the DNA binding region of specific TFs and compensatory evolution in the TFBS.
A special script was developed to test the significance of highly abundant binding signals of specific TFs.
We examined whether the promoter regions of specific genes or the binding sites of specific TFs have undergone more TFBS changes in the lineages leading from wild strains to fermentation or lab strains.
When this number is equal to or larger then a specificity threshold, t, such significant DNA loci (specific BEs) could be used to construct and analyze of statistically reliable part of the empirical frequency distribution representing relatively high-avidity binding loci of specific TF-DNA binding.
One of our most unexpected findings is that epigenetically active and repressed enhancer regions are enriched for the binding sites of two non-overlapping sets of specific TFs.
172– 175; and (3) ChIP-seq on TFs or cofactors (p300, CEBPB, etc).. 176 However, the mapping of open chromatins and histone modifications usually lacks sufficient resolution and specificity to detect precise enhancer locations, and the binding of some specific TFs or cofactors can hardly cover all the active enhancers.
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