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Approximately 90% of all ras mutations affect the K-ras gene, of which 85% occur in codon 12. K-ras mutations occur in particularly high frequency in adenocarcinoma (20 30%) but sequencing of k-ras and c-raf of our transgenic tumor model did not evidence any sequence alteration (data not shown).
However, hematoxylin/eosin staining of liver sections showed no macroscopic signs of tissue alteration (data not shown).
IPA analysis of common DEGs highlighted steroid hormone biosynthesis among deregulated canonical pathways but could not predict any liver function alteration (data not shown).
We were not able to detect any significant differences in gene expression patterns between the NaCl, NMDG-Cl or sucrose methods of tonicity alteration (data not shown).
Similarly, the P53M214K mutant embryos did not show significant vascular alteration (data not shown) and P21 and caspase-8 gene expression was unaltered (Fig. 4H) following genetic depletion of TNFRSF1B.
To analyze the effect of this mutation, we performed CANT1 cDNA analysis on patient 8 RNA extracted from leucocytes and found no product by RT-PCR supporting an absence of CANT1 mRNA transcription due 5′UTR splice site alteration (data not shown).
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The guanidinium chloride insoluble Alcian blue stained Muc2 band showed no alterations (data not shown).
Analysis of the M10 cells-derived metastatic MT10 and MG10 cells revealed the presence of identical gene alterations (data not shown).
The remaining 10 patients displayed H19DMR-GOM but did not show other causative alterations (data not shown).
Minimal cystic dilation was noted in rare glands, in the absence of any gland crowding or other cytologic alterations (data not shown).
We also found four F354L alterations (data not shown) but we did not consider these as missense mutations as this alteration has previously been reported to be a rare polymorphism of the gene (Launonen et al, 2000).
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