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. 2006 Apr 6;25(8):1764–1774. doi: 10.1038/sj.emboj.7601045

Figure 5.

Figure 5

Mre11 complex is required for ATM and ATR-dependent fork restart. (A) DNA replication in restarting extracts. Chromatin was isolated from Mre11 depleted damaging extracts treated with 5 mM caffeine and 500 μM Roscovitine+40 μM APH (lane 1), 55 μM CPT (lanes 2, 4 and 6) or 300 μM MMC (lanes 3, 5 and 7) and then transferred to restarting extracts that were Mre11 depleted (Mre11 dep), Mre11 depleted supplemented with 500 nM of recombinant Mre11/Rad50/Nbs1 complex (Mre11 dep+Mre11/Rad50/Nbs1) or Mre11 depleted supplemented with Flag-ATR and active Flag-ATM proteins (Mre11 dep+ATM* and ATR). Asterisk (*) indicates that ATM has been isolated from irradiated cells (see Supplementary Materials and methods). Replication was monitored for 90 min in the presence of α-32P-labeled dGTP. DNA was then isolated and run on an agarose gel. (B) Quantification of DNA replication in Mre11 depleted restarting extracts. Extracts were supplemented with 40 μM APH (black bars), 55 μM CPT (gray bars) or 300 μM MMC (striped bars) treated chromatin. Graph shown represents a typical experiment.