Anti cd45.1 percp cy5 5 a20

For immunoblots and immunohistochemistry, anti-ST6GAL1 (R and D Biosystems), anti-β-tubulin (Cell Signaling Technology), anti-PF4 (Peprotech 500-P05), anti-IgD (eBioscience 11–26 c), SNA-FITC (Vector labs) were used. For flow cytometry, SNA-FITC (Vector Labs), biotinylated MAL-II (Vector Labs), anti-Gr1-PE/Cy5 (RB6-8C5), anti-CD11b-BV711 (M1/70), anti-CD45.2-PE/Cy7 (104), anti-CD45.1-PerCP/Cy5.5 (A20), anti-Ly6G-APC (1A8), anti-Ter119-BV510 (TER-119), anti-CD41-BV421 (MWReg30), anti-c-kit-APC/Cy7 (2B8), and anti-Sca-1-PE (D7) (all Biolegend) were used.

Cell populations were identified with anti-B220 (CD45R)-PE-Cy7 (RA3-6B2, eBioscience), anti-CD8a-APC (53-6.7, eBioscience), anti-CD4-AF647 (GK1.5, BioLegend), anti-CD45.1-PerCP-Cy5.5 (A20, BioLegend), and anti-Thy1.1 (CD90.1)-APC-Cy7 (OX-7, BioLegend). Cell surface activation marker levels were assessed by staining with anti-CD86-BV650 (GL-1, BioLegend), anti-CD40-APC (1C10, eBioscience), anti-CD69-PerCP-Cy5.5 (H1.2F3, BioLegend), anti-MHC I (H-2Kb)-PE (AF6-88.5.5.3, eBioscience), and anti-MHC II (I-A/I-E)-FITC (M5/114.15.2, eBioscience). All antibodies were used at a 1:500 dilution. Live-dead staining was performed with DAPI (3 μM wash), or propidium iodide (PI; 1:50 dilution). Data collection was performed on a BD LSRFortessa, BD FACSCanto, or BD LSR II cytometer.