Pe anti cd11a

Flow cytometry to characterize peripheral blood neutrophils, monocytes, or BM hematopoietic stem and progenitor cell profiles was performed as previously described (22 (link)). For the ROS measurement, RBCs or lysed WBCs were labeled with APC/Cy7 LIVE/DEAD (Invitrogen, MP34955) and antibodies APC anti-Ter119 (BioLegend, 116212), anti-CD45 (BioLegend, 103128), PE-Cy7 anti-CD11b (BioLegend, 101216), BV421 anti–Gr-1 (BioLegend, 108445), and APC anti-CD115 (BioLegend, 134410) in staining buffer for 20 minutes at 4°C, and then washed once with staining buffer and resuspended in 10 μM H₂ DCFDA (Thermo Fisher Scientific, 88-5930-74) for another 15 minutes at 37°C. For the lipid peroxidation assay, after labeling, the cells were resuspended in 2 μM C11-BODIPY (Thermo Fisher Scientific, D3861) for 30 minutes at 37°C. For circulating leukocyte adhesion marker detection, PE anti-CD11a (BioLegend, 101107) and L-selectin (BioLegend, 104406) were used. Flow cytometry was performed using the LSR Canto or LSRII (Becton Dickinson) and data were analyzed using FlowJo software (Becton Dickinson). Isotype-matched normal IgG was used as the control in each flow cytometry assay.