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. 2009 Mar 12;28(8):1142–1156. doi: 10.1038/emboj.2009.60

Figure 3.

Figure 3

The MRX complex is recruited to forks and promotes recovery during pauses in replication. ChIP using αMyc (9E10) and αHA (F-7, Santa-Cruz) antibodies as described in Figure 2 was performed on (A) Myc-Rad50 and HA-Polα in a wild-type background (JC528), where enrichment at ARS607 (solid bar) and +14 kb from ARS607 (speckled bar) is compared at the indicated time points after release into 0.2 M HU. (B) FACS analysis was performed as described in Cobb et al (2003). Wild-type (JC467) and mre11Δ (JC371) cells were blocked with α-factor for 90 min (G1) and released into S phase with 0.2 M HU for 3 h before cells were washed and resuspended in YPAD to monitor the progression through S phase. Samples are taken at indicated time points (30, 45, 60, 75, 90, 150) after HU removal. (C) Ethidium bromide stained PFGE for samples arrested in G1 with α-factor for 90 min (G1) before treatment with 0.2 M HU for 180 min (0 min time point). HU was removed and cells were washed and released into YPAD media with samples taken at 50, 100 and 150 min. Southern blot analysis was performed to monitor ChrIV replication (lower panel) with quantification of triplicate experiments for Chr IV replication shown in relative units compared with the signal in G1 using Quantity One (Biorad) comparing wild type (JC467) and mre11Δ (JC371).