Fig. 6: BRCA1–BARD1 enhances RAD51-mediated DNA protection.
From: Mechanism of BRCA1–BARD1 function in DNA end resection and DNA protection
a, DNA protection assays with WRN-DNA2-RPA, in the absence or presence of BRCA1–BARD1 and RAD51, performed at 100 mM NaCl. Top, a schematic of the assay. Red asterisks (*) represent the position of the radioactive labels. Middle, quantitation of DNA degradation. Averages shown; error bars, s.e.m.; n = 7 (in the absence of BRCA1–BARD1) and n = 4 (in the presence of BRCA1–BARD1). Bottom, representative assays. b, Representative protection assays with WRN-DNA2-pCtIP-RPA, in the absence or presence of BRCA1–BARD1 and increasing concentration of RAD51. c, Quantitation of DNA protection assays such as shown in b. Averages shown; error bars, s.e.m.; n = 4. The data were normalized to the corresponding reaction without RAD51 (lane 2). Values without normalization are plotted in Extended Data Fig. 9g. Representative protection assays with WRN (10 nM) and DNA2 (10 nM), in presence of pCtIP and BRCA1–BARD1, not shown. d, Quantitation of DNA protection assays such as shown in Extended Data Fig. 9h. Averages shown; error bars, s.e.m.; n = 3. The data were normalized to the corresponding reaction without RAD51 (lane 2). Values without normalization are plotted in Extended Data Fig. 9i. e, Representative protection assays with WRN (25 nM), DNA2 (25 nM), pCtIP (20 nM) and RPA (215 nM), in the absence or presence of BRCA1–BARD1 and either human RAD51 (HsRAD51) or yeast Rad51 (ScRad51). Top, quantitation of DNA degradation. Averages shown; error bars, s.e.m.; n = 5 for reactions containing HsRAD51, in the presence or absence of BRCA1–BARD1 and n = 3 for reactions containing ScRAD51, in the presence or absence of BRCA1–BARD1. ****P ≤ 0.0001, two-tailed t-test. Source data are provided in Supplementary Fig. 3.
