DNA Damage Inducers and Replication Inhibitors

(i) DNA damage inducer and PI3KK inhibitors. Hydroxyurea (HU; #127-07-1; MilliporeSigma) was prepared as a stock solution at 200 mM, according to manufacturer’s instructions. PI3KK inhibitors, KU60019 (#4176; Tocris Bioscience), AZ20 (#S7050; Selleckchem), and NU7441 (#3712; Tocris Bioscience), were dissolved in DMSO at 10 mM. They were applied to cell cultures at 2 days prior to infection at the final concentrations, as we previously reported: HU at 5 mM, KU60019 at 5 μM, AZ20 at 3 μM, and NU7441 at 1 μM (31 (link), 32 (link), 80 (link)).
(ii) In vitro DNA replication inhibitors. We used HAMNO (#S0148, Selleckchem), T2AA (#21921, Cayman Chemical), aphidicolin (#14007, Cayman Chemical), MK886 (#1311, Tocris Bioscience), and PNR7-02 (#2965, Axon Medchem). Stock solutions were prepared in DMSO at 10 mM. The inhibitors were directly added into the reaction of in vitro replication assays at a final concentration of 100 μM for HAMNO, 1 μM for T2AA, 10 μg/μL for aphidicolin, 200 μM for MK886, and 20 μM for PNR7-02 as previously recommended (44 (link)– (link)48 (link)).