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In looking at downstream cell degradation pathways associated with skeletal muscle break-down and Akt signaling, autophagosome flux, as indicated by the expression ratio of LC3 II to I, was increased by PD90 compared to earlier pre-symptomatic timepoints (P < 0.05, Fig. 3B,D).
Consequently, RT-PCR analysis showed that the cobalt-induced Bcl-2, MDR1 and VEGF gene expression was significantly suppressed by zinc (Figure 5FD, compare CoCl2 lanes with CoCl2/ZnCl2 lanes), as also shown by the expression ratio to GAPDH (Figure 5F, lower panel).
As shown in Figure 1B, the ADR-induced p53AIP1-luc activity was significantly impaired by cobalt, while cobalt alone did not affect it, and in vivo analysis of mRNA levels showed that drug-induced upregulation of p53 apoptotic target genes Bax and Puma was strongly impaired by cobalt (Figure 1C), as shown also by the expression ratio to GAPDH.
fold change, defined by the expression ratio of cancer to normal (= cancer/normal) All these genes had false discovery rate <0.001.
Microarray data were ranked by the expression ratio between the geometric mean of the CD24−/low/CD44+ CD24+/CD44+ populations from the three cell lines (Supplementary Table).
The genes are sorted by the expression ratio of those in GRS layer 2 to in GRS layer 5. Note that Kv4.3 and Kv1.4 genes are much more highly expressed in GRS layer 2 than in GRS layer 5 (ratio; 8.65 and 4.24, respectively).
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This program calculates a Perturbation Factor for each gene, which is computed by normalising the expression ratio with information about its topology in the pathway (i.e., number of genes interacting with it and type of interaction) [ 19].
Microarray data were validated by comparing the expression ratio of 14 genes (adhE, clpB, dnaK, hfq, kpdE, marR, minD, nfrB, ompW, soxS, trxC, wzxE, zntA and znuA) with the results of expression determined by real-time PCR (Additional file 3: Figure S1).
fold change, defined by the negative of the expression ratio of normal to cancer (= - (normal/cancer)) All these genes had false discovery rate <0.001.
a M = log of the expression ratio's, calculated as the average expression level observed in the ccpA mutant divided by the average expression ratio observed in the wild-type.
To facilitate a fair comparison of up- and down-regulated genes, fold change was calculated as follows: for genes with an expression ratio ≥1, the fold change is the same as the expression ratio, whereas folds change of genes whose expression ratio is <1 equals the reciprocal of the expression ratio multiplied by −1 [ 54].
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