1. LY induces replication stress and depletes the pool of available RPA2 for binding to DNA. HeLa cells were treated with 0.4% DMSO, 100 nmol/L LY (A), 2,000 nmol/L hydroxyurea (B), or with both LY and hydroxyurea (C).
LY induces replication stress and depletes the pool of available RPA2 for binding to DNA. HeLa cells were treated with 0.4% DMSO, 100 nmol/L LY (A), 2,000 nmol/L hydroxyurea (B), or with both LY and hydroxyurea (C). Plates were fixed at various time points from 0.5 to 9 hours following compound addition and stained for DNA (Hoechst 33342), RPA2, and pH2AX (S139). The relative intensity for each stain was measured on a single cell basis using the Thermo Scientific Cell Insight NXT. S-phase cells were determined by measuring DNA content and used for further analysis. RPA2 staining intensity (x-axis) was plotted versus pH2AX (S139) staining intensity for each condition and time point. The 0.5-hour DMSO control is representative of all DMSO time points and is illustrated as a starting control condition. Regions with increased ratios of pH2AX (S139) to RPA2 are boxed in red and indicate the depletion of RPA2 associated with replication catastrophe.
Constance King et al. Mol Cancer Ther 2015;14:
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