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. 2012 Aug 1;125(15):3529–3534. doi: 10.1242/jcs.105353

Fig. 3.

Fig. 3.

The spatial distribution of 53BP1 and BRCA1 at DSB sites is established rapidly and persists following irradiation. (A) Irradiated cells (2 Gy), detergent pre-extracted and fixed at indicated times following irradiation, immunostained for BRCA1 and 53BP1. Representative projection images of 3D-SIM datasets (n≥8 per group). (B) Projection and 3D reconstructions of 3D-SIM datasets of cell volumes indicated in A. (C) Cells positive for co-enrichment of BRCA1 and 53BP1 in IRIF were processed as in A. (D) Projection and 3D-reconstruction images constructed from 3D-SIM datasets of cell volumes indicated in C. (E) Projection (PRJ) and single Z-slices show 53BP1 exclusion from BRCA1-associated IRIF core. (F) BRCA1 enrichment is associated with decreased 53BP1 IRIF occupancy. Volumes of isosurface-rendered 53BP1 foci from multiple irradiated cells (2 Gy) positive or negative for BRCA1 co-enrichment were analysed per time point; n = 3 per group, scoring >300 foci and >100 foci at 30 minute and 2 hour time points, respectively. Whiskers indicate the 5th to 95th percentile. ***P<0.0001, Mann–Whitney test.