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Furthermore, we analyzed editing levels of the different subunits in mice pretreated for two weeks with celecoxib.
Similarly, editing levels of 5HT2CR sites were much lower in hESCs and their derivatives compared to those observed in adult human brain [17].
Since the function of AMPA glutamatergic receptors is modulated by the mRNA editing of the different subunits, we analyzed editing levels of GluR2 Q/R and R/G sites, GluR3 and GluR4 R/G sites, GluR5 Q/R site and GluR6 I/V, Y/C and Q/R sites in the hippocampus and cortex.
Next, we tested the variability in editing levels of non-recoding sites.
Using Sanger sequencing, editing levels of known conserved targets of A-to-I RNA editing were analyzed from the cerebral cortex of rats used in the RNA-seq: Gabra3, Cyfip2, Kcna1, Flna, and Blcap (which has three editing sites, known as Y/C, Q/R, and K/R), and the newly identified site in Cog3, using primers given in Table 1.
We analyzed editing levels of the GluR- B transcript at the Q/R and the R/G sites, the GluR- 6 transcript at three recoded positions identified as the I/V, Y/C and Q/R sites, the GluR- 5 transcript carrying the Q/R edited site and the Kv1. 1 transcript carrying the I/V edited site.
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We observed also a significant decrease, from 20.7% to 15.4%, in the average editing level of Alu sequences in the spontaneous differentiation (n = 7 edited Alu regions, p<0.016 according to Wilcoxon signed-rank test; Figure 2A).
The editing level of ndhB-1977 was medium (~57%) in both leaf and floral tissues but the editing efficiency of other four newly identified sites was poor (5 17%).
A modest decrease at the editing level of BLCAP was observed during neuronal differentiation (29.2% 23.6%).
No changes by genotype or KA-treatment were found in the editing level of kainate GluR5 Q/R site in the hippocampus or in the cortex (Table S1).
In all mice tested the editing level of GluR2 Q/R site was virtually 100%, with no variations due to either KA injection or genotype (data not shown).
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