Figure 3.

shu1Δ, mms2Δ, and mph1Δ each reduces the levels of X-mols in smc6-P4 cells with double deletions conferring greater reduction. Cells were arrested using nocodazole and then released into YPD medium containing 0.033% MMS. The replication and recombination intermediates at the ARS305 region 60, 120, 180 and 240 min after release were analyzed by 2D gel electrophoresis followed by Southern blotting (see Materials and Methods). (A) Cartoons indicating the position of the probe and the replication structures. (B and D) Results of 2D gel analyses. X-shaped DNA structures are indicated by arrowheads in smc6-P4. FACS analyses are presented to the right of the gel image; and displayed from bottom to top are the profiles for asynchronous cultures, G2 arrested cells, released cultures at 60, 120, 180, and 240 min. Quantification of the 2D gel results in B and D are plotted in C and E, respectively.