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Confluent cell cultures were washed once with PBS, and then incubated for 20 min at room temperature with NucBlue Live ReadyProbes Reagent (#R37605, ThermoFisher Scientific, Waltham, MA, USA) to stain the nuclei, washed with PBS, permeabilized with 0.1% Triton-X 100 for 30 min, fixed at 37 C for 20 min with 4% Paraformaldehyde in PBS, and blocked with 2% BSA in PBS at room temperature for 20 min