Fig. 4

Transcriptional activation (TA) activity of DBD-BRCT protein variants. HEK293 cells were harvested 24 h post-transfection with the DBD-BRCT WT or variant, pGAL4-e1b-Luc (firefly) and phRG-TK (Renilla). Known benign (green) and pathogenic (orange) BRCA1 control variants were included for comparison. The Dual-Luciferase Reporter system was used to quantify TA activity. Activity was measured in triplicates and in three biological replicates. TA activity was normalised by dividing firefly fluorescence signal by Renilla fluorescence signal. The relative TA activity of each investigated variant was expressed as percentage of WT activity (100%). The mean TA activities are shown with error bars depicting the SD. Dashed lines represent estimated thresholds that indicate reduced TA levels similar to the pathogenic (below orange line) or benign control variants (above green line). Footnotes: TA, Transcriptional Activation; WT, Wild Type; EV, Empty Vector