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. 2017 Feb 28;206(1):497–512. doi: 10.1534/genetics.117.199703

Figure 4.

Figure 4

Induction of NDT80 allows Rad51-mediated DSB repair in the dmc1∆ rad17∆ background. Diploids containing either dmc1∆ mek1-as NDT80-IN (NH2278) or dmc1∆ mek1-as NDT80-IN rad17∆ (NH2365) were transferred to Spo medium for 5 hr, and a final concentration of 1 μM 1-NA-PP1 and/or ED was added at the indicated times. Arrows indicate ED addition. DSBs and COs at the HIS4::LEU2 hotspot were detected by XhoI digestion of genomic DNA probed with a 0.6-kb AgeI/BglII fragment from pNH90 (hunter and kleckner 2001). P1 and P2 = parental bands; CO1 and CO2 = CO bands; DSB = DSB fragments. “t” indicates the time that the Mek1-as inhibitor, 1-NA-PP1, was added. (A) Southern blot of NDT80-IN dmc1∆ mek1-as from one of two independent time courses. (B) DSB fragments as a percent of the total DNA is plotted for each timepoint. The averages between two independent experiments are shown with error bars indicating the range. (C) COs for the experiment shown in (A) plotted as in (B). (D) Southern blot of NDT80-IN dmc1∆ mek1-as rad17∆ from one of two independent time courses. (E and F) DSBs and COs, respectively, for the experiment shown in (D). CO, crossover; DSB, double strand break; ED, estradiol; Spo, sporulation medium.