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

Figure 5. MEF2D regulatory regions act as enhancers and candidate outcome variants increase MEF2D PRE1 and PRE2 activity.

Figure 5

Putative MEF2D regulatory regions containing candidate outcome variants were cloned downstream of a MEF2D-promoter-driven luciferase construct (MEF2D promoter in figure). The reference PRE1, PRE2 and rs11264494 constructs contain the most common haplotypes or alleles in Europeans. Alternative alleles of candidate 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 other haplotypes were also generated: PRE1 hap2 (all alternative alleles of PRE1 ref; 35% frequency in Europeans) and hap3 (all alternative alleles of PRE1 ref except rs11264488; 15% frequency in Europeans); and PRE2 hap2 (all alternative alleles of PRE2 ref; 30% frequency in Europeans). 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 replicate experiments performed in OVCAR8 and JAM cells. P-values were determined by two-way ANOVA followed by Dunnett’s multiple comparisons test (*p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001).