Expression of the
3’UTR-variant mutant allele with estrogen withdrawal. A. MCF-7 cells treated with either 100 nM fulvestrant or estrogen for 48-hours, were transfected with dual luciferase reporter plasmids harboring either the non-variant (G-allele, dark grey) or variant (T-allele, light grey) BRCA1 3’UTR. After a 16-hour incubation dual luciferase activities were measured. Renilla luciferase was normalized to firefly luciferase. T-allele expression was calculated to that of the G-allele. Plotted is the mean and standard deviation of at 3 independent experiments. *p < 0.05; error bars represent the mean ± standard deviation. B. Total RNA was isolated from cell lysates (A) by Trizol extraction. RT-qPCR was utilized to access the effects of fulvestrant and estrogen treatment on mRNA expression of estrogen responsive markers (GREB1 and TFF1). The results were normalized to β-Actin mRNA expression. *p < 0.05; error bars represent the mean ± standard deviation.