Fitc anti mouse ly6d

For FACS or flow experiments, single-cell suspensions of cells were resuspended in a 2% FBS solution and then stained with appropriate antibodies at a 1:100 dilution for 60 min at 4 °C in the dark. Antibodies used were APC anti-human/mouse TREM2 (R&D; FAB17291A), APC/Cy7 anti-mouse CD45 (BioLegend; 103116), Alexa700 anti-mouse CD45.2 (BioLegend; 109822), PerCP/Cy5 anti-mouse CD11b (BioLegend; 101228), BV605 anti-mouse Cd11c (BioLegend; 117334), FITC anti-mouse Ly6d (BioLegend; 138606), PE/Cy7 anti-mouse Vcam1 (BioLegend; 105720), BC510 anti-mouse F4/80 (BioLegend; 123135), PE/Cy7 anti-human CD326 (BioLegend; 324222), and PE anti-human VCAM1 (BioLegend; 305805). Before sorting or flow, SytoxBlue (Thermo Fisher; S34857) was used for a viability dye at a 1:1000 dilution. FACS experiments were run on FACSAria II, and flow experiments were run on LSRII instruments, where BD FACSDiva v8.0.1 was used for data collection. Subsequent analysis was done in FlowJo v10.6.

For FACS or flow experiments, single-cell suspensions of cells were resuspended in a 2% FBS solution and then stained with appropriate antibodies at a 1:100 dilution for 60 min at 4 °C in the dark. Antibodies used were FITC anti-mouse Ly6d (BioLegend; 138606) and APC anti-human/mouse Cd49f (BioLegend; 313616). Before sorting or flow, SytoxBlue (Thermo Fisher; S34857) was used for a viability dye at a 1:1000 dilution. FACS experiments were run on FACSAria II, and flow experiments were run on LSRII instruments. Both instruments used the BD FACSDiva 8.0.1 software for data collection. For scRNA-Seq experiments, cells were only stained with SytoxBlue, and live cells were sorted for downstream processing and experimentation. For EdU experiments, we used the Click-iT Plus EdU Alexa Flour 647 Flow Cytometry Assay Kit as per manufacturer instructions (Invitrogen; C10634). For Apotracker^TM experiments, we used the Apotracker^TM Green reagent as per manufacturer instructions (BioLegend; 427402). FlowJo 10.6 was used for data analysis.