Fig. 2.

Meiotic recombination in pCUP1-SHU1 mek1as strain. (A) One-dimensional gel analysis of pCUP1-SHU1 mek1as cells in various conditions. Copper and/or inhibitor were added to aliquots of the same pre-meiotic culture, and recombination intermediates were quantified at the HIS4LEU2 construct and analyzed as described elsewhere (Kim et al., 2010). -IN, absence of 1-NA-PP1; +IN, presence of 1-NA-PP1, Mom, mom species; Dad, dad species; COs, crossover species; DSBs, double-strand breaks; JMs, joint molecules. (B, C) Quantitative analysis of DSBs and crossover products during the same time points shown in (A). Percentage of total DNA in DSBs and COs is plotted as a function of time. (D) MI + MII represent meiotic division progression as estimated by the number of DAPI foci. (E) Representative two-dimensional gels of SEIs and dHJs. Corresponding images of the entire gels are shown in Supplementary Fig. 3. (F, G) Quantitative analysis of joint molecules in pCUP1-SHU1 cells with or without 1-NA-PP1 and copper. DNA event represents DNA species as percent of the total hybridizing signal at specific time points after the transfer to sporulation medium. SEI, single end invasion; IH-dHJ, interhomolog double Holliday Junction; IS-dHJ, intersister double Holliday Junction. (H) Levels of IH-COs and IH-NCOs in the pCUP1-SHU1 mek1as strain. COs and NCOs were assayed with a HIS4LEU2 tester construct containing BamHI (“Mom”) and NgoMIV (“Dad”) sites. IH-COs and IH-NCOs are shown for each time point sample as percentages of the total DNA.