Identification of Macrophage Phenotypes

To identify macrophage phenotypes, a cell suspension with a concentration of 1–5 × 10⁶ cells/mL was prepared. APC-conjugated anti-F4/80 antibodies (eBioscience, San Diego, CA, USA) and PE-conjugated anti-CD206 antibodies (eBioscience, San Diego, CA, USA) were added for staining. The former was used to identify total macrophages, while the latter served as a marker for M2 macrophages [50 (link)]. All antibodies were used at a concentration of 5 μg/mL. After 30 min of incubation on ice, analysis was performed using a BD LSRFortessa flow cytometer (BD Biosciences, San Jose, CA, USA), and data was processed using FlowJo V10 software (Tree Star, Ashland, OR, USA).