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To identify genes harboring cancer-specific promoter methylation, we examined the methylation status of the 23 genes in five to ten pairs of matched primary CRC (PT) and corresponding normal colon (PN) tissues.
We increased the sample numbers to over 25 pairs of primary CRC (PT) and corresponding normal colon (PN) tissues, and to 13 normal colon mucosa tissues from non-cancer patients (NN).
Eight tumor tissues and corresponding normal colon mucosa tissues were used for western blotting analysis.
However, DNA methyltransferase was expressed at increased levels in tumor tissue compared to the corresponding normal colon tissue.
Genomic DNA was extracted from the tumors and corresponding normal colon tissue samples by phenol/chloroform extraction.
One hybridization with high COX-2 tumor expression versus corresponding normal colon mucosa tissue was also performed.
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We examined 98 surgical resected tumors and corresponding adjacent normal colon mucosa from the cohort of patients (48 men, 50 women, mean ± SD age 64 ± 14 years) with primary CRC we reported previously [ 7].
In corresponding normal muscle and colon tissues, there were very weak expressions of PIG-2.
We performed gelatin zymography for the initial characterization of MMP-2 and MMP-9 in tumors and their corresponding normal mucosa from colon and rectal cancer patients.
However, nuclear CBX7 staining was more prevalent in the glandular epithelial cells and stromal lymphoid cells in the corresponding SMs as well as normal colon biopsies from non-cancer patient controls (Additional file 1: Figure S1).
DNA from matched colon cancer tissues and corresponding normal mucosa from CRC patients as well as DNA from colon cancer cell lines underwent bisulfite modification as described previously [24].
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