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Apo-NsrR was prepared by incubating the purified protein with EDTA and ferricyanide in a molar ratio of protein∶EDTA∶ferricyanide (1∶50∶20) at 25°C for 20 minutes [29] before buffer exchanging the protein with buffer A. Loss of cluster was verified by measuring the UV-visible absorbance spectrum of the treated protein, as described below.
The loss of cluster N2 and the dissociation of the hydrophilic arm together preclude the occurrence of the hydrophobic, physiological quinone reduction reaction in subcomplex Iλ. Figure 2 compares Q1 reduction by complex I and subcomplex Iλ in the presence of exogenous phospholipids and in the presence and absence of rotenone.
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Importantly, DNA binding to the hmpA1 and hmpA2 promoters was abolished by NO, demonstrating that NsrR DNA-binding activity is modulated in response to NO. Treatment of the protein with EDTA and ferricyanide [29] resulted in loss of the cluster (as judged from the complete loss of cluster-associated UV visible absorption features, data not shown).
The reduction in transcripts for the IIA subunit cannot explain the degree of loss of clustering we see at the neuromuscular junction, arguing the problem in the SMN depleted tissue is not primarily based on transcript reduction.
Study arms were balanced at baseline, and there was no loss of clusters to follow-up.
This included recruitment bias, baseline imbalance, loss of clusters, incorrect analysis or selective outcome reporting.
To allow for dropout of individual participants and loss of clusters, we aimed at including 60 clusters.
Moreover, despite a loss of clustering, telomeres remained associated with the NE in RNAi mutants (Hall et al, 2003).
13 They also had appropriate randomisation, comparability of clusters, accounting for cluster effect, and no loss of clusters to follow-up (table 1).
For cluster-RCTs, we addressed additional components: recruitment bias; baseline imbalance; loss of clusters; incorrect analysis; compatibility with RCTs randomized by individual.
Thus, BDNF priming induces β1-integrin clustering that can resist negative remodeling by MAG exposure, but secondary BDNF treatment cannot overcome the MAG-induced loss of clustering.
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