Anti ki 67 16a8

Cells were incubated with anti-CD16/32 (clone: 93) (eBioscience, CA, USA) for 30 min and surface stained with a panel of antibodies at 4°C for 1 h. The following monoclonal antibodies were used in this study: anti-CD11b (M1/70), anti-F4/80 (BM8), and anti-CD169 (SER-4) were purchased from eBioscience, anti-CD45 (clone104) and anti-CD31 (clone390) were purchased from BioLegend (CA, USA), and anti-CD204 (REA148) was purchased from Miltenyi Biotec (Bergisch Gladbach, Germany). For intracellular CD206 staining, the cells were fixed and permeabilized following intracellular staining with anti-CD206 (C068C2, BioLegend, CA, USA) at 4°C for 1 h. For the staining of Ki-67, the cells were stained for surface markers, then fixed and permeabilized for intracellular anti-Ki-67 (16A8, BioLegend) staining using fixation and permeabilization buffers (eBioscience) according to the manufacturer's instructions. For ROS measurement, the cells were incubated with 10 μM Dichloro-dihydro-fluorescein diacetate (DCFH-DA) (Sigma-Aldrich, MO, USA) at 37°C for 20 min. All FACS data were acquired on a FACSCalibur (BD Biosciences) or AriaIII (BD Biosciences) flow cytometer and analyzed using FCS Express V3 and FlowJo 7.6 software.