Fig. 1: Microtubule-dependent nuclear deformations and DSBs mobility.
a Immunoblot for BRCA1, CHK2 and phospho-CHK2 (P-CHK2) in p53-deleted (sg-P53) Brca1F/F Mouse Embryonic Fibroblasts (MEFs) 72 h after transduction with Hit&Run Cre and/or 6 h treatment with PARPi (0.5 μM). Actin is shown as loading control. b Examples of 10 min traces of mCherry-BP1-2 foci in Brca1F/F MEFs 72 h after Brca1 deletion, 6 h after PARPi addition in the absence or presence of taxol (1 h) or nocodazole (2 h). Scale bars: 10 μm. c MSD of mCherry-BP1-2 foci in the indicated MEFs as described in (b), with SD. Total foci analyzed: 1618 for DMSO, 1056 for taxol, and 1085 for nocodazole from 35, 30, 28 nuclei, respectively from n = 3 independent experiment. d Representative image of Brca1F/F MEFs without any treatment or 72 h after Cre-mediated deletion of Brca1, PARPi treatment for 6 h treatment and/or incubation with taxol for 1 h. Highlighted boxes indicate nuclear deformations as vertical invaginations (blue), light (green) and deep (yellow) lateral invaginations, and longitudinal invaginations (magenta). In black and gray boxes, the control nuclear edges and nuclear interior with no invaginations, respectively. Scale bars: 2 μm. Magnification ×4.25. (e) Quantification of nuclear deformations as in (d) from 40, 20 and 22 cells for each condition derived from one representative experiment. Statistical analysis by Kruskal–Wallis test for multiple comparisons. (*) P < 0.05 (p = 0.0381); P ≧ 0.05 are not significant. Source data are provided as a Source Data file. See also Supplementary Fig. 1.
