Fixed cells were then incubated with blocking solution containing the DNA dye FxCycle Violet (1 μg/mL, Thermo Fisher #F10347) for at least 1 hour at RT in the dark before samples were run using the BD Biosciences LSRFortessa™ flow cytometer and analysed using FlowJo® and GraphPad Prism software.

Fixed cells were then washed once with 1xPBS before being incubated in 500 μL blocking buffer (1xPBS +2% BSA + 0.1% TritonX-100/Tween20) for 1 hour at room temperature (RT), primary antibody solution (phospho-MPM-2, 1:500, Merck Millipore #05-368 phospho-histone H2AX S139, 1:500, Merck Millipore #05-636) in blocking buffer for 2 hours at RT and secondary antibody solution (Alexa Fluor 488 Goat anti-mouse, 1:500, Thermo Fisher #A11017) in blocking buffer for 1 hour at RT in the dark with three washes using blocking buffer between each step. The Lancet Regional Health – Western PacificĬells were collected via trypsinisation at the indicated times, washed with 1xPBS and fixed in ice-cold 70% ethanol overnight at -20☌.The Lancet Regional Health – Southeast Asia.The Lancet Gastroenterology & Hepatology.

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