Avatar
Gymsushi5

0 Following 0 Followers
1
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
1
Nhibitor TSA on the mRNA expression of MAGE-A1 gene and the other family members (MAGE-A2, -A3 and -A12) in different cell lines. Moreover, we assessed the methylation status of the MAGE-A promoters by sodium bisulfite mappingbefore and after stimulation with the demethylating agent 5-aza-CdR and/or TSA. While the methylation patterns clearly correlated with the basal MAGE RNA transcript levels, u
1
Lyzed its impact on the modifications of histones bound at the MAGE-A1 promoters. To investigate the changes in the histone signature of MAGE-A1 promoter, it was compared in basal MCF-7 cells (no expression of MAGE-A1, Table 1) to the signature in MCF-7 cells stimulated by 5-aza-CdR with/without TSA or transfected with the expression plasmid encoding for BORIS. For these analyses we used antibodie
1
Lyzed its impact on the modifications of histones bound at the MAGE-A1 promoters. To investigate the changes in the histone signature of MAGE-A1 promoter, it was compared in basal MCF-7 cells (no expression of MAGE-A1, Table 1) to the signature in MCF-7 cells stimulated by 5-aza-CdR with/without TSA or transfected with the expression plasmid encoding for BORIS. For these analyses we used antibodie
1
Lyzed its impact on the modifications of histones bound at the MAGE-A1 promoters. To investigate the changes in the histone signature of MAGE-A1 promoter, it was compared in basal MCF-7 cells (no expression of MAGE-A1, Table 1) to the signature in MCF-7 cells stimulated by 5-aza-CdR with/without TSA or transfected with the expression plasmid encoding for BORIS. For these analyses we used antibodie
1
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
1
Rexpression of Ets-1 could not further upregulate the promoter activity mediated by BORIS, as shown by several repetitions. Surprisingly, co-transfection with an expression plasmid encoding for Sp1 partly repressed the stimulatory effect mediated by BORIS (p = 0.001), whereas the addition of expression plasmid encoding for Ets-1 abrogated this repression (Figure 5B). To verify the repressive effec
1
Rexpression of Ets-1 could not further upregulate the promoter activity mediated by BORIS, as shown by several repetitions. Surprisingly, co-transfection with an expression plasmid encoding for Sp1 partly repressed the stimulatory effect mediated by BORIS (p = 0.001), whereas the addition of expression plasmid encoding for Ets-1 abrogated this repression (Figure 5B). To verify the repressive effec