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
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
L line BCM1, the expression of both genes is opposite: no levels of MAGE-A1 [2^(Ct) 1.07] and high levelsof BORIS [2^(Ct) 24.39]. We transiently transfected expression plasmid encoding BORIS into both cell lines, with negligible transcript levels of MAGE-A1, and quantified endogenous MAGE-A1 mRNA by RT (reverse transcription)-PCR and gel electrophoresis. As depicted in Figure 1, BORIS was able to
Ults show that changes in the BORIS transcript levels are associated with those of MAGE-A1 and corroborate that BORIS is involved in the activation of MAGE-A1 gene expression.BORIS affects the DNA methylation pattern of MAGE-A1 geneFigure 2 BORIS mRNA expression in MDA-MB-468, MCF-7 and BCM1 cells, untreated or treated with 5-aza-CdR. RT-PCR products of BORIS mRNA were separated on an agarose gel.
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
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
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
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