Fig 1: Validation of microarray results by real-time PCR and Western blot. A. Comparison of cDNA microarray expression ratios (Tca/PYM-Con expression over Tca/PYM-siTCRP1 expression) with those determined by real-time PCR for seven genes. The relative fold changes were calculated after normalizing against ß-actin expression. Ratios of transcripts in Tca/PYM-Con and Tca/PYM-siTCRP1 cells were comparable to those obtained in the microarray analysis. Each column represents the results of three independent experiments. B. We further validated the expression of CYP3A5, HIF1A, TP53, MT1X, NF?B1, Akt1, and EGFR proteins in both cell lines, using ß-actin as a loading control to investigate the correspondence between mRNA and protein levels for these different genes. Similar changes as those found in the microarray analysis were observed. Each figure represents three independent experiments (a: TcaPYM-siTCRP1 cells, b: Tca/PYM-Con cells).