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Expression of these mutant constructs showed that mutations of L857 to Y and S exhibited similar effect on cell cell fusion as the L857-F mutant.
Analysis of various TSP1 promoter mutant constructs showed that a sequence located −144/−137 up-stream of the transcriptional initiation site, related to the consensus E2F-responsive sequence, is necessary for the activation.
Furthermore, flies rescued by the wildtype and two mutant constructs showed similar developmental timing, gauged by pupation curves (Figure 2C), and similar final animal size, measured by wing area (Figure 2D).
In contrast, the percentage of eGFP-positive cells after transfection with both mutant constructs showed a significant increase at 24 hours and 36 hours.
In contrast, myotubes transfected with both mutant constructs showed no increase in 'chymotrypsin-like' enzyme activity in the presence of PIF.
At the protein level, as shown in Figure 3D, the seven mutant constructs showed generally lower expression than the construct g-a-c (−15.8~−51.1%) and among them, constructs a-a-t, a-c-c, a-a-c, g-c-c, and a-c-t reached the significance level of 0.05, with their UGDH protein concentration being reduced by 37.8%,51.1%, 34.7%, 26.5%, and 50.1%, respectively.
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While the data for R53A binding was uninterpretable due to severe line-broadening of NMR signals, studies with the R57A, R56A, and R52A mutant constructs show that interactions of individual arginines occur independently of the neighboring ASM (Supplementary Fig. 8f).
One-dimensional H NMR spectra recorded for the mutant constructs show the cytosine substitutions do not affect the global secondary structure of SL3ESS3 as diagnostic imino signals of base-paired regions were observed.
This −216/−1 mutant construct showed promoter activity equivalent to that of the wildtype −216/−1 region, indicating that basal MUC4 expression in T47D cells was not dependant on the Ets binding site at position −216.
Chick retinas electroporated with CR4.2-Hand-mutant construct showed no change in GFP expression as compared to CR4.2-GFP expression (Fig. 5E G), while transfection of CR4.2-mut-Meis1-βGP-GFP construct diminished GFP expression (Fig. 5H J).
In contrast, the mutant BAP1 constructs showed reduced or no inhibitory effects according to the possible deterioration effects of the mutants (Fig. 3a).
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