Cd49d pe clone r1 2

Blood samples were collected from the retro-orbital plexus of anesthetized mice in sodium heparinized capillaries at a 50:50 ratio of blood to 1% dextran plus 10 U/ml sodium heparin. After erythrocyte sedimentation, the overlying supernatant plasma-dextran suspension of leukocytes was washed in phosphate-buffered saline (PBS), followed by erythrocyte lysis. The leukocytes were then labeled with the following anti-mouse antibodies: anti-CD45-Vioblue (clone REA737, Miltenyi Biotec), anti-CD11b-APC-Cy7 (clone M1/70, Biolegend), anti-Ly6G-FITC (clone REA526, Miltenyi), anti-CD8α-PE-Cy7 (clone 53-6.7, Biolegend), anti-CD4-APC (clone RM4-4, Biolegend), cd49d-PE (clone R1-2, Biolegend). Cell viability was detected using 7AAD (eBioscience). Specimens were acquired by flow cytometry using a MACSQuant Analyzer (Miltenyi Biotec). Data were analyzed using FlowJo software.

CD3-FITC clone 145–2C11, CD4-PE-Cy7 clone RM4–5, CD4-PE clone RM4–4, CD11a-FITC clone M17/4, CD49d-PE clone R1–2, CXCR5-biotin clone L138D7, PE-Cy7 streptavidin, CD44-FITC and CD44-APC-Cy7 clone IM7, PD-1 APC clone 29F.1A12, GL7-PacBlue and GL7-PE clone GL7, CD19-PerCP-Cy5.5 clone 6B2, CD138-BV421 clone 281–2, IgD-APC-Cy7 clone 11–26c2a, CD95-PE clone SA367H8, CD73-BV605 clone TY/11.8, CD71-BV421 Clone RI7217, and CD80-PE clone 90, were purchased from Biolegend (San Diego, CA). BcL6-PE-CF594 clone K112–91 was purchased from BD Biosciences, San Jose, CA)