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In addition, degradation tags (LVA) or LacI binding sites were tested for their effects and possible efficacy at reducing background leaky expression (data not shown).
Since these binding sites were tested in vivo, evidence of sequence conservation was sought, as described.
Moreover, in cultured hepatocytes that express Foxa2 endogenously, ectopic expression of Insm1 and Neurod1 can synergistically stimulate transcriptional activity in luciferase assays when fragments containing Insm1/Neurod1/Foxa2 binding sites were tested.
Randomly selected binding sites were tested for sequence and verified by ChIP coupled with real time PCR, demonstrating the existence of a GATA3 binding site and binding affinities corresponding to relative peak intensities (data not shown).
Binding sites were tested against 20 false clusters of genes (genes selected randomly from those represented on the Affymetrix Drosophila Genome 1 array and treated the same as true clusters).
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The reversible operation of the photocontrolled binding sites was tested with alternating UV and visible light illumination.
The pΔ(A25 -GFP construct feA25 -GFPdeletion of the constructSp1 binding sites, was tested featuringence upon expression.
In Mab21l2 and Tgfb2 promoter regions, twelve putative individual Pax6-binding sites were tested by electrophoretic mobility shift assays (EMSAs) with recombinant Pax6 proteins.
In addition, all SNPs in haploblock 1 in the 5' regulatory region (spanning over 17 kb, chr 3: 173648897–17365897; NCBI accession number for GHSR NM_198407.1) available in the HapMap database and located in putative TF-binding sites were tested for their functionality in gelshift experiments.
A 560 bp fragment spanning both putative BRK sites was tested for binding to immobilized BRK protein and binding was demonstrated.
Although one study used conventional APLs in collagen-induced arthritis [ 33], unconventional APLs with substitutions at MHC binding sites were mainly tested in arthritis models.
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