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Our definition of presence of a gene in a given condition requires that it expresses more than the median expression intensity.
Therefore, we have used median expression intensity cutoff as a measure to categorise genes as expressed or not expressed [ 3].
The median expression intensity for all P2 DAlFEs was slightly lower than the P1 counterparts (22.3 versus 27.1 respectively; see Table 1 for a summary by tissue), but was not significantly different.
A gene was therefore scored for its presence (or absence) depending on the median expression intensity in each condition (Methods).
Genes were functionally grouped (Hes6, Hes6-Chga, Hes6-Ddc, Ascl1, Ascl1-Hes6-Nts-Ddc) and a median expression intensity calculated per group for each sample.
Total 18 expression data obtained by microarray analysis were exported to GeneSpring GX (Agilent Technologies) and per chip normalization to the quantile expression level and per gene normalization to the median expression intensity were performed in all samples.
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Median CEBPA expression intensity was 148 (range: 21 960).
The median CEBPA expression intensity value in all patients was 670 (range 48 5244).
Read counts were further normalized for each gene's expression intensity (median read count) to avoid expression differences becoming unrecogniseable due to outliers.
As almost all of the carcinomas showed a high percentage of cells expressing VEGF (median expression 93%), we defined high expression as intensity of staining >2 based upon the median intensity of staining.
Median expression values for the three intensity values were calculated, disregarding values exceeding 3.5-fold difference from the median.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com