Anti cd44 mab

Icariin was purchased from MEYER company (Shanghai, China), whereas the antibodies used for western blotting were purchased from Cell Signaling Technology (Danvers, MA). The antibodies were anti-CD3 mAb (PE), anti-CD4 mAb (APC), anti-CD8a mAb (PE/Cy7), anti-CD25 mAb (APC/Cy7), anti-CD44 mAb (FITC), anti-CD62L mAb (APC/Cy7), anti-IFN-γ mAb (PE), HO-1 mAb (PE/Cy7), anti-CD16/32 mAb and anti-Foxp3 mAb (FITC) (eBioscience, San Jose, CA, USA).

Cells were lifted with Accutase (ThermoFisher Scientific, Franklin, MA, USA) and washed in phosphate‐buffered saline pH7.4 and then stained with Fixable Viability Dye eFluor 450 and antibody against Sca‐1 (eBioscience, Franklin, MA, USA). For indicated experiments, cells were further stained with anti‐CD44 mAb (eBioscience) using standard immunofluorescence staining method. The Sca‐1 single‐ or Sca‐1⁺CD44⁺ double‐positive cells were quantified by FACS (Gallios, Beckman Coulter, Pasadena, CA, USA) and analyzed with FlowJo or FACScan. Dead cells were omitted from marker quantification based upon viability dye staining. To examine Sca‐1 expression by ICC, cells (1 × 10⁴ per plate) were cultured in small culture plates for indicated days after radiation treatment, cytocentrifuged onto the slide, and then stained with anti‐Sca‐1 antibody by standard ICC staining method and examined under light microscope. For Fig. S1A, cells were cultured on coverslips, fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton‐×100. Cells were then stained with Sca‐1 antibody and mounted using ProLong Gold Antifade Mountant with DAPI (ThermoFisher Scientific).