Pe anti mouse f4 80 bm8

CD1 mice were subcutaneously injected in their hock regions with DiD-labeled α7-NTs, and their draining lymph nodes were collected at 6 or 24 h after injection for imaging and quantification using a PerkinElmer Xenogen IVIS 200 imaging system. For cellular level analysis, single-cell suspensions were generated from lymph node samples by manual dissociation, passage through Flowmi 40-μm cell strainers (Bel-Art), and 2 washes with PBS. The cell suspensions were then incubated with a LIVE/DEAD fixable aqua dead cell stain kit (Invitrogen), blocked with 1 % BSA in PBS, and stained with a panel of antibodies, including FITC anti-mouse CD3 (17A2, BioLegend), Pacific Blue anti-mouse CD19 (6D5, BioLegend), PE/cyanine7 anti-mouse CD11c (N418, BioLegend), PE anti-mouse F4/80 (BM8, BioLegend), APC/cyanine7 anti-mouse CD11b (M1/70, BioLegend), and PerCP anti-mouse Ly-6G/Ly-6C (Gr-1) (RB6-8C5, BioLegend). B cells were defined as CD19⁺, macrophages were defined as CD11b⁺F4/80⁺, dendritic cells were defined as CD11c⁺F4/80^-, granulocytes were defined as CD11b⁺Gr-1⁺, and T cells were defined as CD3⁺. Unstained, single-stained controls, and fluorescence-minus-one controls were used for compensation and gating purposes. Data were acquired using a Becton Dickinson LSR II flow cytometer and analyzed using FlowJo v10.4 software.