Fig. 7: Overexpression of EXO1 rescues overcomes Arp2/3 inhibition.
A Resection analysis of FZ201 (pGal:exo1). Samples were collected at 0, 1, 2, 3, 4, 5, and 6 h after adding galactose. Resection analysis was done from a minimum of 3 biological replicates collected in triplicate. See “Methods” section for more details. Mean ± SEM is shown. Source data are provided as a Source Data file. B Diagram of the experimental setup to measure the MSD of a DSB labeled with Ddc2-GFP. CK-666 (100 µM) was added 2 h 40 min after adding galactose. C MSD analysis (∆t = 30 s) of DSB relative to the SPB in FZ015 (n = 15), FZ201 (pGal:EXO1) (n = 14), and FZ201 with CK-666 (100 µM) (pGal:EXO1) (n = 13) 3 h after Gal-HO induction. CK-666 (100 µM) was added 20 min before image collection as described in Fig. 1C. Imaging and analysis were done as described in (B). Mean ± SEM is shown. Source data are provided as a Source Data file. D RC from MSD analysis of strains in (C). 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). WT vs. pGal::EXO1 (p = 0.6748), WT vs. pGal::EXO1 + CK-666 (p = 0.4505), and pGal::EXO1 vs. pGal::EXO1 + CK-666 (p = 0.0737). See Supplementary Data 1 and 2 for significance and RC values. Mean ± SEM is shown. Source data are provided as a Source Data file. E Resection analysis of FZ015 (WT) and FZ218 (Las17-AID pGal::EXO1). IAA was added either 1 h before galactose or 1 h after galactose. Samples were collected at 0, 1, 3, and 6 h after adding galactose. Resection analysis was done from a minimum of 3 biological replicates collected in triplicate. See “Methods” section for more details. Mean ± SEM is shown. Source data are provided as a Source Data file. F MSD analysis (∆t = 30 s) of DSB relative to the SPB in FZ015 (n = 10), FZ218 (pGal:EXO1 Las17-AID) (n = 10), and FZ218 with IAA (1 mM) 2 h after galactose (pGal:EXO1 Las17-AID + IAA) (n = 10). 3 h after Gal-HO induction. CK-666 (100 µM) was added 20 min before image collection as described in Fig. 1C. Imaging and analysis done as described in Fig. 1C. Mean ± SEM is shown. Source data are provided as a Source Data file. G RC from MSD analysis of strains in (F). 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). WT vs. pGal:EXO1 Las17-AID (p = 09371), pGal:EXO1 Las17-AID vs. pGal:EXO1 Las17-AID + IAA (p = 0.6209), and WT vs. pGal:EXO1 Las17-AID + IAA (p = 0.9172). See Supplementary Data 1 and 2 for significance and RC values. Mean ± SEM is shown. Source data are provided as a Source Data file. H Western blot of Las17-AID pGal::EXO1 probed with α-Myc to show Las17-AID degradation and phosphorylated protein, and α-Pgk1 as a loading control. Source data are provided as a Source Data file. I Western blot of Las17-AID pGal::EXO1 with IAA added 1 h before DSB induction, probed with α-Myc to show Las17-AID degradation and α-Pgk1 as a loading control. Source data are provided as a Source Data file. J Western blot of Las17-AID pGal::EXO1 with IAA added 2 h after DSB induction, probed with α-Myc to show Las17-AID degradation and α-Pgk1 as a loading control. Source data are provided as a Source Data file.
