Fig. 2: Arp2/3 is required for DSB-induced chromatin mobility.
A GFP-tagged chromosome locus in FZ152. LacO-array is 4.4 kb away from a Gal-HO inducible DSB in the MAT locus on chromosome III labeled with GFP-LacI. SPB is labeled with Spc42-mCherry. B MSD analysis (∆t = 30 s) of the MAT locus relative to the SPB before Gal-HO induction (uncut, 0 h) (n = 12), treatment with CK-666 (100 µM) before Gal-HO induction (uncut + CK-666, 0 h) (n = 10), and 3 h after Gal-HO induction (cut, 3 h) (n = 15). Imaging and analysis done as described in Fig. 1C. Mean ± SEM is shown. Source data are provided as a Source Data file. C Experimental setup to measure the MSD of a GFP-LacI/lacO array 4.4 kb away from the HO-cut site on chromosome III. CK-666 was added 20 min before adding galactose, and MSD was measured 3 h after DSB induction (CK-666 before Gal-H), or CK-666 was added 2 h 40 min after galactose and MSD was measured 3 h after DSB induction (CK-666 after Gal-HO). D MSD analysis (∆t = 30 s) of the MAT locus relative to the SPB before Gal-HO induction (uncut, 0 h) (n = 12), 3 h after Gal-HO induction where CK-666 (100 µM) was added 20 min before galactose (cut + CK-666 before gal, 3 h) (n = 10), and 3 h after Gal-HO induction (cut, 3 h) (n = 15). Imaging and analysis done as described in Fig. 1C. Mean ± SEM is shown. Cut and uncut data from (B) is shown again as a comparison for CK-666 treatment. Source data are provided as a Source Data file. E MSD analysis (∆t = 30 s) of the MAT locus relative to the SPB before Gal-HO induction (uncut, 0 h) (n = 12), 3 h after Gal-HO induction with CK-666 (100 µM) (cut + CK-666 after gal, 3 h) (n = 17), and 3 h after Gal-HO induction (cut, 3 h) (n = 15). Cells treated with CK-666 were given CK-666 20 min before imaging. Imaging and analysis done as described in Fig. 1C. Mean ± SEM is shown. Cut and uncut data from (B) is shown again as a comparison for CK-666 treatment. Source data are provided as a Source Data file. F Rc from MSD analysis of strains in (B), (D), and (E). Statistical analysis for the radius of confinement derived in PRISM using a one-way Anova (ns p ≥ 0.05, *p < 0.05, **p < 0.01, and ***p < 0.001). 0 h vs. 0 h + CK-666 (p = 0.18), 0 h vs. 3 h (p = 0.0012), 0 h vs. 3 h pre-CK-666 (p = 0.6039), and 0 h vs. 3 h post-CK-666 (p > 0.999). See Supplementary Data 1, 2 for significance and RC values. Mean ± SEM is shown. Source data are provided as a Source Data file.
