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. 2016 Feb 1;7(9):9759–9772. doi: 10.18632/oncotarget.7112

Figure 2. rs7250266 and rs2278256 reduce NBA1 promoter activity.

Figure 2

(A) Illustration of luciferase reporter constructs containing rs7250266 or rs2278256. DNA fragment of NBA1 promoter region (−964 to +308 bp, the transcriptional start site is determined as +1) containing rs7250266 (C > G), rs2278256 (T > C) or both was cloned into pGL3 luciferase vector respectively. (B) Luciferase reporter assay of cells expressing constructs containing different alleles of rs7250266 and rs2278256. MCF-10A, MDA-MB-468 or MDA-MB-231 cells were transiently transfected with luciferase reporter constructs as illustrated and the internal pRL-CMV control plasmid. Luciferase activity from cells carrying constructs containing the C-allele of rs7250266 and T-allele of rs2278256 (pGL3 −620C/−73T) were set to “1” and used for control for comparison. Fold change was calculated by comparing luciferase activity from cells expressing other alleles of rs7250266 and rs2278256 to cells expressing pGL3 −620C/−73T. Data represent mean values, with error bars indicating S.E.M. (standard error of the mean). Statistical data were analyzed by the t test (*P < 0.05; **P < 0.01). Experiments have been repeated three times for each cell line and similar results were obtained.