Extended Data Fig. 2: 53BP1-MD relation to underlying chromatin.
From: Stabilization of chromatin topology safeguards genome integrity
a, 3D-SIM of GFP–53BP1-MDs in U2OS cells exposed to irradiation (1 Gy, 2 h) and immunostained for γH2AX (PCC = 0.93, n = 300 MDs) shows high colocalization of 53BP1 and γH2AX. b, STED of a γH2AX-MD in U2OS cells treated as in a. c, 3D-SIM of three different z-planes of HeLa cells expressing histone H2B–GFP, treated with 10 ng ml–1 NCS for 2 h and immunostained for γH2AX. Nuclear DNA was visualized by DAPI. Insets are magnified γH2AX-MDs. Intensity line profiles of the three fluorophores (along the white line in the insets) show colocalization of chromatin with γH2AX-MDs. d, Western blotting of U2OS cells treated with control or XRCC4 siRNA immunostained for XRCC4 and loading marker (KAP1). e, f, Intensity line profiles of 53BP1-MDs with XRCC4 (e) and RPA (f) in cells treated as in Fig. 1b; six independent examples per condition are shown. Fluorescence intensities in c, e, f were normalized to the maximum value of each profile. Scale bars, 200 nm in a, b, insets in c and 5 μm in whole-nucleus images (c). Experiments were biologically replicated twice with similar results. For detailed image information see Supplementary Table 1. For gel source data see Supplementary Fig. 1.
