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Many mutants were active in one pathway, but not the other, demonstrating that the two activities of IRF3 are not inter-dependent.
As we had shown that all the NPHP-disease mutants were active in HEK 293 cells, we tested whether there was any change in activation of these proteins upon serum starvation in hTERT-RPE1 cells.
These two deletion mutants were active in the disintegration assay in the presence of Mn2+ conditions (Table I), consistent with previous results [21], [23], [24].
Most MT2 mutants were active in the cAMP assay with the exception of MT2-R138C and to a lesser extend the MT2-R231H MT2-K243R243R mutants (Figure 4A).
All of the T4 Rnl2tr mutants were active in strand-joining reactions except the R55K K225R ligase.
All the mutants were active; however, major changes in the kcat values were measured (15- to 500-fold decrease).
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Having established that all the disease mutants are active and that activity is key to localization, we re-investigated the localization of these mutants.
This, along with he fact that mutants are active and the enzyme turns over 800 times before the crosslink is formed, suggests that the thioether bond is not a crucial structural requirement for cysteine oxygenation.
However, the finding that a p73-PxxP mutant is active in transcription activation but not in apoptosis, argues that transcription activation might be required, but not sufficient, in mediating apoptosis.57 Interestingly, a similar mechanism was proposed for p53.
In this study we found that the ΔF508/V510D mutant was active while we failed to detect activity when V510D was introduced into Cys-less CFTR (9).
Analysis of the activity of CDK9's S175A and S175D mutants showed that CDK9 S175A mutant was active and S175D - inactive.
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