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. 2017 Jun 15;8(39):64670–64684. doi: 10.18632/oncotarget.18501

Figure 2. ZNF100 PRE1 and PRE2 enhance promoter activity in OVCAR8 cells.

Figure 2

ZNF100 PRE1 and PRE2 regions containing the major alleles of candidate outcome variants were cloned downstream of a ZNF100-promoter-driven luciferase construct (ZNF100 promoter in figure) for the creation of reference (PRE1 ref and PRE2 ref) constructs. Minor allelic variants of the outcome variants were engineered into constructs and are designated by the rs ID of the corresponding variant and a black box in the construct schematic. Constructs containing haplotypic variants occurring in Europeans were also generated: PRE1 hap2 (minor allelic variants of all candidate variants except for rs4808295; 10% frequency in Europeans) and PRE1 hap3 (all minor allelic variants; 4% frequency in Europeans); and PRE2 hap2 (all minor allelic variants). Cells were transiently transfected with each of these constructs and assayed for luciferase activity after 24 h. Back-transformed data are shown and error bars denote 95% confidence intervals of experiments performed in triplicate for OVCAR8 cells and in quadruplicate for COV362 cells. P-values were determined by two-way ANOVA followed by Dunnett’s multiple comparisons test (*p < 0.05, **p < 0.01 and ***p < 0.001).