Sensitivity of <i>BARD1</i> variants to cisplatin.

(A) Cells expressing BARD1 WT, A460T, P707S, G753D, and V767fs cells, as well as endogenous-only cells, were treated with control, BARD1 3'UTR or BRCA1 3’UTR siRNA and treated with cisplatin at concentrations of 1.875, 3.75, 7.5, and 15 μM. Colonies were counted after 12 days of growth followed by staining with crystal violet. Results in each experiment were done in triplicate and converted to logarithmic scale (±S.E.M.). In each plot, the dashed line indicates control siRNA, and the solid line indicates depletion with the BARD1 or BRCA1 3'UTR-targeted siRNA. The Student’s t-test was done to examine the growth of variant and endogenous-only cell lines treated with BARD1 or BRCA1 siRNA relative to variant and endogenous-only cells treated with control siRNA (indicated by asterisks; * = p BARD1 or BRCA1 3'UTR siRNA. A Student’s t-test was done to compare the colony formation of BARD1 variant cell lines, as well as endogenous-only cells depleted of BRCA1 or BARD1, to BARD1 WT cells. Colony counts from variants and endogenous-only cells were significantly different (p 0.05). (C) An examination of endogenous BARD1 and BRCA1 expression in treated BARD1-expressing and control cell lines. Cells treated with BARD1 3'UTR siRNA had depleted endogenous BARD1 expression, while HBT BARD1 was unaffected. Cells treated with BRCA1 3’UTR siRNA showed depletion of endogenous BRCA1 and BARD1 expression. The band representing the expression of variant HBT BARD1 was denser than the endogenous BARD1 protein band in all cases.