Cell cycle analysis using in vivo EdU labeling

EdU (Thermo Fisher Scientific) was dissolved in sterile 1× PBS or DMSO, and 1 mg in a volume of 100 μl was injected i.p. 1 h before sacrifice. EdU detection was performed with the Click-iT Plus EdU Pacific Blue and Alexa Fluor 647 Flow Cytometry Assay Kits (Thermo Fisher Scientific). Cells were stained for surface markers for 15 min at 4°C, followed by fixation and incubation with the Click-iT Plus reaction cocktail according to the manufacturer’s instructions.