A De Novo Noncoding RARB Variant Associated with Complex Microphthalmia Alters a Putative Regulatory Element

Figure 4

Assessing the effect of the c.157+1895G>A variant on the transcriptional regulation of RARB. (a) Schematic of luciferase (luc) reporter design. RARB promoter and the conserved region 1 (CR1), WT and variant (carrying c.157+1895G>A), were cloned into a basic pGL4.10 [Luc2] vector and transfected into human lens epithelial cells or HEK293 cells; chr3 coordinates for each RARB region are indicated based on hg19. Resultant fold change in luciferase activities from transfected (b) human lens epithelial or (c) HEK293 cells showed significantly increased activity in the presence of the CR1 harboring the c.157+1895G>A variant. Data were derived from three independent replicates, each performed in triplicate. One-way ANOVA with Tukey’s post hoc test was used to determine the statistical significance. . ns = nonsignificant. Lines represent mean fold response. Error bars represent standard deviation.

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