Hmgb1 antibody

During 0-21 days of tumor inoculation, the above tumor-bearing mouse were injected with 100 μL PBS (Hyclone), 20 μg/mL Mouse IgG2b, κ Isotype (Biolegend), 5 or20 μg/mL HMGB1 antibody (Biolegend) every 3 days for total seven times, respectively. And the subcutaneous tumor sizes were measured every 3 days. At day 21, the mice were sacrificed, spleen and tumor tissues were extracted, and the single cell suspension was prepared. The levels of CD3, B220, CD11c, CD11b, and Gr-1 in spleens and the levels of CD11b, Gr-1, F4/80, CD4 and Foxp3 in tumors were detected using flow cytometry.

The cell concentration of sorted lineage-depleted bone marrow cells was adjusted at 1×10⁶/mL, and the cells were added into a 24-well plate (1 mL/well,20 wells are used). The wells with cells were randomly divided into five groups: phosphate-buffered saline (PBS) (Hyclone), 10 μg/mL HMGB1 (Biolegend), 10 μg/mL Mouse IgG2b, κ Isotype (Biolegend), and 2 and 10 μg/mL HMGB1 antibody (Biolegend) were addeded, respectively. All cells were incubated with 20 ng/mL IL-6 (R&D) and GM-CSF (R&D) for 5 days, and the levels of CD3, B220, CD11c, CD11b and Gr-1 were detected using flow cytometry.