F4 80 viogreen

Macrophages from peritoneal exudate derived from mice in the TGC model (0.5 × 10⁶ cells) were resuspended in FACS buffer (PBS supplemented with 0.5% bovine serum albumin and 0.1% NaN₃) and were labeled with the antibody mixes from panel 1 (CD45-APC; F4/80-Viogreen; MHCII-Vioblue; LFA-1-PeVio770; CD64-PE; CD16/32-Viobright FITC) (Miltenyi Biotec, Bergisch Gladbach, Germany) and panel 2 (CD45-APC; F4/80-Viogreen; CD64-PE; MHCII-Vioblue; CD38-PeVio770; CD206-FITC) (Miltenyi Biotec, Bergisch Gladbach, Germany) for 20 min at 4°C. Cells were then washed with FACS buffer and the relative fluorescence intensities were determined using a MACSQuant Vyb cytometer equipped with FlowLogic software (Miltenyi Biotec, Bergisch Gladbach, Germany).