Cd11c fitc

To detect the impact of aAGd-NWs+RT on specific antigen presentation, B16-OVA subcutaneous tumor model was established on C57 mice. Initially, 5 × 10⁵ B16-OVA cells were subcutaneously injected into the mice. After a period of 7 days, when the average tumor size in the mice carrying B16-OVA tumors reached approximately 100 mm³, the mice were grouped randomly into six distinct categories: Vehicle, GGd-NCPs, aAGd-NWs, Vehicle+RT, GGd-NCPs+RT, and aAGd-NWs+RT ([Gd] = 15.7 mg kg⁻¹ and [ara-AMP] = 34.7 mg kg⁻¹). Treatments were performed on day 0 and day 6, respectively, and followed by RT (5 Gy × 2) 6 h post treatment. On day 8, tumor draining lymph nodes were harvested and processed into single-cell suspensions. The cells thus obtained were stained for 30 min using a combination of FITC-CD11c and APC-H2Kb-SIINFEKL antibodies (BioLegend, USA). Subsequently, these cells were analyzed through flow cytometry using the BD FACSCalibur system (USA).

Cell suspensions were stained with appropriate antibodies for 30 min on ice. The commercial antibodies used in this study included anti-mouse FITC-CD45 (BioLegend, CA, USA, #103108; 1 µg per 10⁶ cells), PE-CLEC2 (BioLegend, CA, USA, #146104; 1 µg per 10⁶ cells), BV421-F4/80 (BioLegend, CA, USA, #123137; 1 µg/10⁶ cells), Pe/Cy7-CD64 (BioLegend, CA, USA, #128016; 1 µg per 10⁶ cells), Percp/Cy5.5-CD11b (BioLegend, CA, USA, #101228; 1 µg per 10⁶ cells), APC/Cy7-PirB (R&D, CA, USA, #FAB2754S; 5 µg per 10⁶ cells), APC-Ly6C (BioLegend, CA, USA, #128016; 1 µg per 10⁶ cells), APC-Ly6G/Ly-6C (Gr-1) (BioLegend, CA, USA, #108411; 1 µg per 10⁶ cells), PerCP-CD19 (BioLegend, CA, USA, #115531; 1 µg per 10⁶ cells), APC-CD3 (BioLegend, CA, USA, #100235; 1 µg per 10⁶ cells), APC-CD206 (BioLegend, CA, USA, #141707; 2 µg per 10⁶ cells), and FITC-CD11c BioLegend, CA, USA, #117305; 1 µg per 10⁶ cells); and anti-human PE-CD14 (BD Biosciences Pharmingen, USA, #555398; 20 µl per 10⁶ cells), Mouse anti-human LILRB2 (R&D, R&D Systems, MN, USA, #MAB2078; 0.25 µg per 106 cells), and Goat Anti-Mouse FITC-IgG (Servicebio, Wuhan, China, #SF131; 1:200 dilution). All antibodies were diluted according to the manual from the manufacturer’s website. Dead cells and doublets were removed by dead-cell dye staining (Zombie Aqua Fixable Viability Kit, BioLegend, CA, USA, #B297827).