Fig. 4

Effect of novel splicing variants on transcription/translation activity in cells. a Schematic diagram depicting how the transcriptional and translational activity were measured. Firefly luciferase expression vectors with each splicing variant and Renilla luciferase expression vector were transfected to HeLa cells followed by DNA, mRNA, and protein extraction after 48 h. The relative expression level of firefly luciferase mRNA was measured through quantitative PCR normalized with the house-keeping gene human RPS18 and the transfection efficiency by genomic luciferase and human BRCA2 exon 27 region from DNA samples. To determine the efficiency of translation activity, firefly and Renilla luciferase activity were detected using dual-luciferase reporter assay normalized by transcriptional activity. b Relative transcriptional efficiency of canine BRCA2 splicing variants containing luciferase vector. c Relative translational efficiency of canine BRCA2 splicing variants containing luciferase vector. d Relative transcriptional efficiency of human BRCA2 splicing variants containing luciferase vector. c Relative translational efficiency of human BRCA2 splicing variants containing luciferase vector. “**” indicates p < 0.01