Cd206

Mouse lungs were fixed in 10% neutral-buffered formalin. Sections (5 μm) were stained for H&E and immunohistochemistry (IHC). Antibodies against F4/80 (1: 250; Cell Signaling Technology) and CD206 (1:100; Abcam) were used for IHC analysis. For analysis of BALF, cells were stained for eosinophils, macrophages, neutrophils, and lymphocytes using flow cytometry. Eosinophils were defined as SSC^high SiglecF⁺ Mac-3⁻ cells, alveolar macrophages were identified as SSC^high SiglecF⁺ Mac-3⁺ cells, granulocytes were recognized as SSC^high Gr-1⁺ cells, and lymphocytes were identified as FSC^low/SSC^low cells expressing CD3 or CD19.

Immunohistochemistry was performed according to the protocol previously described 22 (link). Briefly, the brain tissue was removed from the anti-freezing solution, and the immunohistochemistry procedures were followed. The primary antibodies used were those targeting arginase (ARG; 1:50; Santa Cruz, CA), CD206 (1:200; Abcam, Cambridge, MA), Iba-1 (1:200 dilution, WAKO, Osaka, Japan), mitogen-activated protein 2 (MAP2; 1:200, 1:100; Millipore, Billerica, MA) and NeuN (1:200, Millipore). TdT-mediated dUTP nick end labeling (TUNEL) and NeuN double staining was performed following the manufacturer's instructions on double immunostaining (Roche Diagnostics, Basel, Switzerland). We performed statistical analysis of the immunohistochemistry data by counting the number of CD206 and IBA1, ARG and IBA1, CD16/32 and IBA1, TUNEL and DAPI double-positive cells and GFAP positive cells in 5 regions in the striatum of peri-infarct area of each brain slice, with 2 brain sections (600 μm apart) in each mouse, 4 mice per group.