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. 2016 Mar 23;7(18):25885–25901. doi: 10.18632/oncotarget.8292

Figure 6. Influence of combinations of checkpoint abrogators on the focalization of BRCA1 and Rad51 induced by trabectedin or lurbinectedin.

Figure 6

A. HeLa cells were exposed to 10 nM trabectedin (left panel, T) or lurbinectedin (right panel, L) for 1 hour in the absence (white columns) or presence of 2 μM KU-60019 (+ KU, light grey columns), 1 μM VE-821 (+ VE, medium grey columns) or a combination of 2 μM KU-600019 and 1 μM VE-821 (+ KU + VE, dark grey columns). This was followed by 24 hours post-incubation in the absence (white columns) or presence of 2 μM KU-60019 (+ KU, light grey columns), 1 μM VE-821 (+ VE, medium grey columns) or a combination of 2 μM KU-600019 and 1 μM VE-821 (+ KU + VE, dark grey columns). Cells were then pre-permeabilized with ice-cold CSK-lysis buffer, fixed and immunolabeled with a BRCA1-directed antibody. Untreated cells were used as a negative control (black columns). The fluorescence intensities in single cells were quantified by Metamorph analysis and are expressed in arbitrary units (a.u.). Data are represented as mean +/− SD. B. (trabectedin) and C. (lurbinectedin), Same as above, except that cells were directly fixed and immunolabeled with a Rad51-directed antibody. DNA was counterstained with Topro-3 fluorescent dye. Rad51 focalization was visualized by confocal microscopy.