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. Author manuscript; available in PMC: 2024 Feb 15.
Published in final edited form as: Mol Cell. 2023 Jul 26;83(16):2925–2940.e8. doi: 10.1016/j.molcel.2023.06.031

Figure 3. BRCA2 nucleates RAD51 at the edges of ssDNA gap in the presence of RPA.

Figure 3

(A) Schematic of 5.374 knt-long ssDNA embedded within λ dsDNA.

(B) Positional analysis of RAD51(A647) (red) binding along the length of 5.374 knt ssDNA gap. 200 nt bins. n = 5 molecules. Error bars represent SEM. 200 nt bins.

(C) Position analysis of RAD51(A647) (red) or BRCA2-eGFP (blue) binding along the length of 5.374 knt ssDNA gap in the absence of RPA. n = 6 molecules. Error bars represent SEM. 200 nt bins.

(D) Position analysis of RAD51(A647) (red) or BRCA2-eGFP (blue) binding along the length of 5.374 knt ssDNA gap in the presence of 1.25 nM RPA. n = 11 molecules. Error bars represent SEM. 200 nt bins.

(E) Per-bin normalized A647 signal frequency for 3’ and 5’ ds-ssDNA junction (3 and 3 bins) or middle of the ssDNA gap (21 bins) in the indicated conditions. Error bars represent SEM. p values by Student’s t test. n.s., p > 0.05; *p ≤ 0.05; **p ≤ 0.01. See also Figure S2.