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. 2015 Jul 8;43(15):7371–7387. doi: 10.1093/nar/gkv691

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© The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 6. — EXO1 modifies the cellular response to low dose camptothecin. (A) Measurement of replication fork tracts upon CPT treatment in the indicated genotypes. The tract length ratios (IdU/CldU) were not significantly different with or without 80 nM CPT treatment in the initial 20 min. The genotypes are abbreviated in each case as follows: wild type = W, Nbs1^ΔB/ΔB = N, Exo1^−/− = E, and Nbs1^ΔB/ΔB Exo^−/− = NE. (B) Analysis of BrdU incorporation at the indicated times post CPT addition by flow cytometry. The percentage of cells incorporating BrdU drops more dramatically in Nbs1^ΔB/ΔB cultures after 24 hours of treatment compared to other genotypes. Time points (0, 5 and 24 h post CPT treatment) were assessed (legend corresponds to panels B and C). (C) Cell cycle profiles of cell cultures of the indicated genotypes following CPT treatment and withdrawal. (D) Metaphase aberrations induced by 50 nM CPT treatment and 4 h of recovery in the indicated genotypes. (E) Sensitivity of cell cultures of the indicated genotypes transfected with siRNA (siGFP as a control or 2 siRNAs against Mus81) to the indicated dose of camptothecin using the colony formation assay. Average results from triplicate experiments with 2 different Mus81 siRNAs (n = 6) are plotted and standard deviation indicated. (F) Average fold increase in survival following the depletion of MUS81 calculated by dividing the fraction surviving values of the MUS81 siRNA treated cells by the siGFP controls for each of the genotypes.