Anti ly6g alexafluor488

Single‐cell spleen suspensions were prepared at 0, 4, 5, 6, 7, 8, 9, 10, 14, 17, 21, 24 and 28 days post‐infection (dpi) as previously described.13 Unless otherwise stated, cell suspensions were re‐suspended in 0.05% FBS BD FACSFlow Sheath Fluid. Cell washings were carried out by centrifugation at 314 g for 7 minutes. Incubations were performed at 4°C for 30 minutes. Non‐specific binding sites were blocked using anti‐CD16/CD32 (Fc γ III/II block—final dilution 1/1000). Afterwards, 5 × 10⁵ cells were incubated with antibody cocktails (dilution of 1/600), using anti‐B220‐FITC, anti‐CD1d‐PE, anti‐CD138‐PE/CY7, anti‐CD93‐APC, anti‐CD4‐FITC, anti‐CD8a‐PE, anti‐TCR β chain‐APC, anti‐Ly6G‐AlexaFluor488, anti‐Ly6C‐PE, anti‐CD11b‐APC, anti‐NK1.1‐APC and anti‐Ter119‐PE (BioLegend, San Diego, CA, USA), 1 µg of 7‐amino‐actinomycin D (7AAD) to exclude nonviable cells, and finally analysed using a BD Accuri™ C6 Plus flow cytometer.