Sc0227

Sox9^CreER, Rosa^tdTomato and Rosa^DTA mice were obtained from The Jackson Laboratory. For lineage tracing, Sox9^CreER; Rosa^tdTomato mice were intraperitoneally injected with tamoxifen (Sigma, T5648-1G) at a dose of 0.08 mg/g body weight every day for three consecutive days. After radiation treatment, Sox9^CreER, Rosa^tdTomato and Rosa^DTA mice were sacrificed at different time points (3, 7, 14 and 30 days). For PI3K/AKT inhibitor treatment, irradiated Sox9^CreER; Rosa^tdTomato mice were assigned to the control and perifosine (250 mg/kg body weight per week, Beyotime, SC0227) treatment groups. For PI3K/AKT agonist treatment, irradiated Sox9^CreER; Rosa^tdTomato mice were assigned to the control and SC79 (5 mg/kg body weight per week, Beyotime, SF2730) treatment groups. After 3, 7 and 14 days of treatment, lung samples were collected and injected with EdU 2 h prior to euthanasia. Both male and female mice were used, and all animal experiments were performed according to the Institutional Animal Care and Use Committee-approved protocols. For euthanasia, the mice were placed in new cages, euthanized immediately with 100% carbon dioxide and decapitated within 5 min for tissue collection.