Upregulate MAGE-A1 expression in this cell line. BORIS-specific shRNA reduced the BORIS mRNA expression nearly completely in presence and absence of scramble shRNA (Figure 3D, p = 0.0001). Likewise, the downregulation of MAGE-A1 expression by BORIS-specific shRNA was more prominent in MCF-7 cells than in MDA-MB-468 cells. As measured by quantitative real time PCR, BORIS-specific shRNA reduced the
Ontrol. At 48 or 72 hour post-transfection, we quantified the changes in the BORISSchwarzenbach et al. BMC Cancer 2014, 14:796 http://www.biomedcentral.com/1471-2407/14/Page 7 ofAMAGE-A1 429 bp _ ?Actin 202 bp _MCF-BMAGE-A1 429 bp _ ?Actin 202 bp _BCMMAGE-A1 mRNA Expression ( )C80000 60000 40000 20000 2000 1500 1000 500MCF-p=0.p=0.p=0.001 p=0.basalAZATSAAZA + TSABorisFigure 1 Comparison of the MAG
Nhibitor TSA or after transient transfection of these cells with an expression plasmid encoding for BORIS were separated on an agarose gel. The bar chart shows the relative changes in mRNA expression levels of MAGE-A1 in MCF-7 cells by quantitative real-time PCR. The significant p-values are shown (C). H2O lane serves as a negative control. The housekeeping gene -Actin was selected as an internal