Pe anti mouse siinfekl h 2kb antibody

MC38.OVA tumour-bearing mice were injected intravenously with PBS, IMDQ, NPGN and PGN_4.9 nanoadjuvants (equivalent to 2 mg kg^-1 IMDQ) every four days for three times. Four days after the last treatment, popliteal lymph nodes and inguinal lymph nodes were dissected and prepared into a single-cell suspension. The cells were stained with Brilliant Violet 510 anti-mouse F4/80 (123135, Biolegend, clone number: BM8, Dilution 1:40), PE-Cy7 anti-mouse CD80 antibody (104734, Biolegend, clone number: 16-10A1, Dilution 1:40), APC anti-mouse CD86 antibody (105012, Biolegend, clone number: GL-1, Dilution 1:80), and PE anti-mouse SIINFEKL-H-2K^b antibody (116608, Biolegend, clone number: SF1-1.1, Dilution 1:40) and detected by flow cytometry. Meanwhile, tumour tissues were harvested and digested by a tumour dissociation kit (Miltenyi Biotec). The single-cell suspension in different groups was stained with PerCP/Cy5.5 anti-mouse CD45 (103132, Biolegend, clone number: 30-F11, Dilution 1:100), FITC anti-mouse/human CD11b (101205, Biolegend, clone number: M1/70, Dilution 1:200), Brilliant Violet 510 anti-mouse F4/80 (123135, Biolegend, clone number: BM8, Dilution 1:40), and PE anti-mouse SIINFEKL-H-2K^b antibody (116608, Biolegend, clone number: SF1-1.1, Dilution 1:40). SIINFEKL⁺ cells in CD11b⁺F4/80⁺ macrophages were measured by flow cytometry.

BMDMs were treated with various IMDQ formulations, and OVA_257-264 (SIINFEKL, 10 μg mL⁻¹, Sangon Biotech) or OVA_257-280 (SIINFEKLTEWTSSNVMEERKIKV, 10 μg mL⁻¹, Biomatik) for 24 h. Antigen-presenting cells were collected for staining of costimulatory factors, including PE-Cy7 anti-mouse CD80 antibody (104734, Biolegend, clone number: 16-10A1, Dilution 1:40), APC anti-mouse CD86 antibody (105012, Biolegend, clone number: GL-1, Dilution 1:80), as well as PE anti-mouse SIINFEKL-H-2K^b antibody (116608, Biolegend, clone number: SF1-1.1, Dilution 1:40) to evaluate the antigen-processing capacity of BMDMs by flow cytometry.