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Overexpression of the PUM2 mutant which fails to interact with Aurora-A, and depletion of PUM2 result in a decrease in the amount of Aurora-A.
The CIITA L1035P mutant, which fails to translocate to the nucleus [58], demonstrated increased Gag processing and virus release compared to vector-only control, suggesting a cytoplasmic role for CIITA in the later stages of the HIV infection cycle.
Interestingly, the PI3Kγ K251E mutant, which fails to bind to Ras [22] and hence to translocate to the endosome via activated Ras (Figure S1A, B), was unable to display a compensatory effect, unlike its wild-type counterpart (Figure 1A), while the expression levels were shown to be comparable by immunoblotting (Figure S1C).
We isolated an Arabidopsis mutant which fails to induce [Ca2+]cyt elevation in response to exudate preparations from various microbes.
Here we describe an A. thaliana mutant which fails to induce [Ca2+]cyt elevation in Arabidopsis roots in response to the exudate preparations from pathogenic root-interacting microbes.
To address whether other UNC93B1-dependent TLRs require AP-2 versus AP-4 to reach endosomes, we examined whether signaling is affected in macrophages expressing the UNC93B1-Y539A mutant, which fails to recruit AP-2.
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The G39A mutant, which failed to discriminate between specific and non-specific RNA binding, was also inactive in DNA demethylation as well as activation of EGFP expression.
Overexpression of the PUM2 mutant, which failed to interact with Aurora-A, and depletion of PUM2 both led to the destabilization of Aurora-A (Figure 3).
Notably the K45A mutant, which failed to discriminate between specific and non-specific RNA binding, was fully active in the demethylation assay, indicating that the two properties can be uncoupled.
To provide insights into the specificity, we also tested the U1F9spec together a scrambled AON on the +5T mutant, which failed to rescue splicing (Supplementary Material, Fig. S3B).
Therefore, we reasoned that expression of CIITA mutants which fail to localize to the nucleus should not mediate increased Gag processing or enhanced infectious virus release.
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