Figure 6

Characterization of MCF-7 cells stably expressing DCD. Quantitative RT-PCR (A) and western blot (B) analysis confirming the expression of DCD mRNA and protein in MCF-7-DCD cells. DCD transcript levels were normalized to that of HPRT using REST software and the expression level in MCF-7 pcDNA control was designated as 1. C, Quantitative analysis of colony numbers (>25 cells/colony) two weeks after plating of the cells. D, Cellular survival of control and DCD overexpressing MCF-7 cells treated with H₂O₂, at indicated concentration, for 16 hrs. Cell survival rates were estimated by MTT assay. Y-axis indicates % surviving cells as percentage of untreated control. Results are given as means ± SD of three experiments. * p < 0,01 and ** p < 0,001 by Student t test. Quantitative RT-PCR analyses showing the fold-increases in the mRNA expression of ligands (E) and the receptors (F) of ERBB family in DCD expressing MCF-7 cells compared to control. Y-axis indicates mean ± SD mRNA levels (normalized to HPRT) relative to controls. G, Effects of EGF treatment on the phosphorylation of EGFR and downstream signaling proteins p38-MAPK and pAkt in MCF-7 control and DCD-expressing cells following the treatment with EGF 10 ng/ml for 15 min.