Flow Cytometry Analysis of Peripheral Blood

For the flow cytometry analysis of the peripheral blood cell detection, we used the strategy recommended by Liu et al. [18 (link)]. The samples were preincubated with anti-mouse CD16/CD32 (Miltenyi Biotec, Bergisch Gladbach, Germany, 130-092-574) for 15 min. Then the following antibodies (all from Miltenyi Biotec) were used: anti-mouse Ly6G–VioBlue (130-119-902), anti-mouse FcεR1–PE (130-118-896), anti-mouse SiglecF–PE-Vio615 (130-112-330), anti-mouse CD172–PE-Vio770 (130-123-154), anti-mouse CD45–APC (130-110-798), anti-mouse CD11b–APC-Vio770 (130-113-803). The antibodies were used in dilution 1:30. The samples were incubated for 30 min and washed twice with DPBS. SytoxGreen (Invitrogen, Eugene, OR, USA, S34859) was added (in dilution 1:1,000,000) 5 min before the acquisition. The measurement was performed using a MACSQuant Analyzer 10 (Miltenyi Biotec).