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Gene expression analyzes were performed using 3 arrays, representing 3 biological replicates with one replicate done in a dye-swap manner to test for dye-based bias.
These morphological and anatomical data were supported by our gene expression analyzes.
Previous gene expression analyzes (also using Affymetrix expression arrays) in myeloma cells have identified DUSP6 as one of only three genes which were uniquely and strongly elevated in cells harboring activated N-RAS [ 28].
After RNA digestion with 1U RNase A (USB, Cleveland, USA) for 1 h at 37°C, gene expression analyzes were performed by adding 0.1 μL of the cDNA to FluoCycleTM sybr green (20 μl final volume; Euroclone, Pero, Italy) in a DNA Engine Opticon Real-Time PCR Detection (Biorad, Hercules, USA).
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During a typical day I spend time doing "gene expression," analyzing what distinguishes the genetic signature of a normal person from a diseased person.
Total RNA was then isolated and gene expression analyzed using Clontech Atlas Rat 1.2 Toxicology arrays.
Figure S4 shows clusters of gene expression analyzed using DAVID.
RNA samples were prepared and gene expression analyzed as described previously [7].
We therefore tested whether ERK signaling in T cells also induces hexokinase gene expression, analyzing the same mRNA samples that we used previously to study glutaminase expression.
Differential gene expression, analyzed with Illumina Beadstation, was used to detect intracellular signaling resulting in changed transcription, compared to control cells (n = 2 in each group).
The chips were scanned on the Illumina Bead Array Reader (Illumina, San Diego, CA) and gene expression analyzed.
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