Figure 5: Induction of apoptosis as a result of T3 Mediated repression of hSMP 30 in contrast to absence of any significant induction of apoptosis as result of E2 mediated repression of hSMP 30. (A,B) A representative qRT-PCR data for two apoptotic related genes BCL-2 and BAX expression in MCF-7 and MDA MB-231 cells. Shown are the mean of triplicate samples (mean SD) normalized to GAPDH. CT values obtained from the real time PCR was used to compare the expression label of treated sample from control assuming 100% amplification. Results were confirmed in three independent experiments. P<0.0001 difference from only vector transfected control using ANOVA for BCL-2 gene, P<0.0232 difference from control using ANOVA for BAX gene expression in MCF-7 cells. P<0.0001 difference from only vector transfected control using ANOVA for BCL-2 gene, P<0.001 difference from control using ANOVA for BAX gene expression in MDA MB-231 cells. Over expression of SMP30 gene after transfection has been confirmed by RT-PCR. SMP30 gene in (upper panel) and GAPDH gene in (lower panel) are shown using cDNA from MCF-7 and MDA MB-231 cells. (C,D) Apoptosis was assessed post vector/SMP30 cDNA transfection and E2/T3 treatment by flow cytometry. Shown are the mean of triplicate samples SD normalized to control. Similar findings were found in two other experiments, P < 0.001 difference from only vector transfected control using ANOVA in Figure 5C and P < 0.01 difference from only vector transfected control using ANOVA in Figure 5D.