Cd11a pe cy7

For primary monocytes, 20 µl of citrate blood was diluted in 80 µl PBS for staining within 4 h after blood collection. Primary monocytes and THP-1 cells were stained with following antibodies (diluted 1:50): CD45-PE (Cat: 368510), CD14-FITC (Cat: 367116), CD16-BV-510 (Cat: 302048), HLA-DR-APC-Cy7 (Cat: 307618), CX3CR1-BV421 (Cat: 341620), CD11a-PE-Cy7 (Cat: 301220), CD11b-PerCP (Cat: 101230), CD11c-BV421 (Cat: 371512), CD29-PE-CY7 (Cat: 303026), and CD49d-APC (Cat: 304308) (all from Biolegend, San Diego, USA). After 25 min staining in the dark, 650 µl RBC Lysis Buffer (Biolegend) were added to the samples and incubated for another 20 min. Subsequently, suspension was centrifuged at 400 × g for 5 min and supernatant was discarded. Cell pellet was resuspended in 100 µl fresh PBS and used for FACS analysis (Supplementary Figure 1S). Flow cytometry was performed with a MACSQuant 10 flow cytometer (Miltenyi Biotec, Bergisch-Gladbach, Germany) and data were analyzed using the FlowJo software version 10.0 (FlowJo, LLC, Ashland, USA).

Collection and preparation were previously described (12 (link)). Red blood cells were lysed using ice-cold deionized water for 30 s. White blood cells were then resuspended in FACs buffer [1× phosphate buffered saline (PBS), 2% heat-inactivated fetal bovine serum (FBS)], counted, and blocked for 10 min in FACS Buffer and Human Trustain FcX Fc receptor blocking solution (Biolegend; #422302) at room temperature, followed by staining with CD3-APC/Fire750 (BioLegend; #344840), CD56-PE (BioLegend; #318306), CD8-PerCP (BioLegend; #344708), CD11a-PE/Cy7 (BioLegend; #301220), CD16-FITC (BioLegend; #302006), CD69-APC (BioLegend; #310910). The corresponding immunoglobulin G (IgG) isotype controls were used for staining the lymphocytes. Cells were analyzed on a BD FACSCanto™ II, and the FACs data were analyzed using FlowJo V.10 data analysis software.

Within 4 h after blood collection, 20 µl of citrate blood was diluted in 80 µl PBS. Blood cells were stained with the following Abs: CD45-PE, CD14-FITC, CD16-BV-510, HLA-DR-allophycocyanin-Cy7, CX3CR1-BV421, CD11a-PE-Cy7, CD11b-PerCP, CD11c-BV421, CD18-allophycocyanin, CD162-PE, CD29-PE-CY7, and CD49d-allophycocyanin (all from BioLegend, San Diego, CA). After 25-min staining in the dark, 650 µl RBC Lysis Buffer (BioLegend) was added to the samples and incubated for another 20 min. Subsequently, the suspension was centrifuged at 400 × g for 5 min, and supernatant was discarded. Cell pellet was resuspended in 100 µl fresh PBS and used for FACS analysis.