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In an effort to characterize the transcriptome of prion-infected murine tissues, we have searched for transcripts which (1) are profoundly upregulated and (2) whose predicted gene products contain secretory leader peptides.
Based on our dataset encompassing expression in 22 different murine tissues, we identified 845 genes with predicted 3'UTR extensions.
To examine the relative expression of Mup transcript(s) in adipose tissues vs. other murine tissues, we conducted qPCR and Northern blot analysis.
Given the lack of available Rxrα ChIP-seq data in murine tissues, we determined the proportion of Rxrα binding sites that coincided with diverse epigenetic modifications identified by the Mouse ENCODE Consortium in liver, brain (by combining cortex and cerebellum data sets) and small intestine [ 17].
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To detect IFN-γ levels in murine colon tissues, we resected approximately 30 mg colon tissues from control and experimental mice which we homogenized in 150 μl NP-40 lysis buffer and centrifuged at 14, 000 g for 30 min.
Nnat expression is particularly low in the murine tissues in which we found Blcap_v1a to be biallelically expressed (Fig. 2B).
Furthermore, in murine pancreatic tissue we identified cells expressing αSMA and the C5a receptor, suggesting that PSCs express the C5a receptor on their surface.
Using this approach, we characterized the extracellular matrices of normal murine tissues (e.g., lung and colon) and demonstrated that each of these comprises over 100 proteins.
We used a microarray dataset consisting of 22 different murine tissues, with 3 5 replicates for each tissue (in total 70 microarrays).
We show that BbCRASP-2 is ubiquitously expressed in diverse murine tissues, but not in ticks, reinforcing a role of BbCRASP-2 in conferring B. burgdorferi defense against persistent host immune threats, such as complement.
Based on the sequences published in GenBank and Ensembl, we designed specific primers and detected by PCR three mRNA species in murine tissues and eight variants in human cells.
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