Fig 1: BRCA2 and PALB2 localization at the centromeres(A) Schematic of the experimental protocol for IAA depletion and staining using a Halo ligand (Janelia Fluor 549 [JF549]). Shown is the volume overlap analysis criterion.(B) Representative images of mAID-Halo-BRCA2 and PALB2-mAID-Halo, visualized using JF549 Halo ligand, CENP-B by IF, and DNA stained by Hoechst. The scale bar equals 5 μm. The inset scale bar equals 1 μm.(C) Number of Halo foci representing the fusion proteins detected using the 3D segmentation pipeline. Each datapoint represents one nucleus, and colored dots are means of the replicates. Paired t test. n = 3. Error bars represent SEM.(D) Number of CENP-B foci detected using the 3D segmentation pipeline. Each datapoint represents one nucleus, and colored dots are means of the replicates. Error bars represent SEM. Paired t test. n = 3.(E) Percentage of CENP-B or RAD51 focus volume overlapping with Halo-BRCA2 and PALB2-Halo foci. Values are the relative frequency of overlap from all colocalization events from the non-treated condition. RAD51 data are from Figure S4E.(F) Percentage of nuclei with none or at least one colocalization event between CENP-B and Halo. 3 biological replicates with 100 cells analyzed in each condition per replicate. Error bars represent SEM.(G) Number of colocalization events per nuclei. Colored dots represent the means of the replicates. Error bars represent SEM. Paired t test. ns represents no statistical significance; p < 0.05 (∗), p < 0.01 (∗∗), p < 0.001 (∗∗∗).See also Figures S2, S3, and S4.