Abstract
<p>PDF - 68KB, Protocol for Microarray analysis and Quantitative RT-PCR (qPCR) MM.1S, U266, and 8266R5 cells were treated with 20, 40, and 40 microM PRIMA-1Met, respectively for 8 hrs and total RNA was isolated. Gene expression was analyzed with Illumina RNA analysis Beadchips (Illumina Inc. San Diego, CA) representing ∼48,000 human genes (Human HT12) as described by us earlier. (28.29) Array data analysis was carried out with Bead Studio software as reported previously. (28,29) Genes showing at least a 2.0-fold difference in expression levels between control and PRIMA-1Met-treated cells were considered to be modulated by PRIMA-1Met. To quantify and validate the expression of p53 target genes of interest at their mRNA level, qRT-PCR assays using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a reference gene were performed as described previously. (28,29)</p>
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