Figure 3

Validation of genechip results by Real-time PCR and Western blotting. A: Using Real-time PCR, we validated the expression of MAP2K6, RECQL, CCND1, CCND3, ABCB1, ABCC2 and GST-Pi transcripts in both cell lines and calculated the relative fold changes by normalizing against GAPDH expression. Ratios of transcripts in Tca/cisplatin cells and Tca8113 cells generally showed the same expression differences as microarray analysis. Each column represents the results of an independent experiment (repeated three times). B: Using Western blotting, we further validated the expression of CCND1, CCND3, ABCB1 and GST-Pi proteins in both cell lines, using β-actin as a loading control. The results also show alterations similar to those found using the genechip. Each figure represents three independent experiments. (a: Tca/cisplatin cells, b:Tca8113 cells).