Dnase

FSBs of silver carp were purchased from a local aquatic product market (Shanghai, China). Sodium dodecyl sulfate (SDS) was purchased from Shanghai Macklin Biochemical Co. (Shanghai, China). Triton X-100 was obtained from Shanghai Titan Scientific Co. (Shanghai, China). DNase and RNase were purchased from Shanghai Yuanye Bio-Technology Co. (Shanghai, China). EGCG, PC, PCA and TA were obtained from Shanghai Macklin Biochemical Co. (Shanghai, China). The cell proliferation and cytotoxicity assay kit (CCK-8 kit) and fetal bovine serum (FBS) were obtained from Beyotime Biotech (Shanghai, China). The aforementioned chemicals were used as received without further purification.

MC38 or B16-OVA tumor–bearing C57BL/6 mice were euthanized, and tumor tissues were harvested and digested with 10 U/mL collagenase I, 400 U/mL collagenase IV, and 30 U/mL DNase (Yuanye Bio-Technology) to generate single-cell suspensions. The following fluorescently labeled antibodies were then used for staining of different markers: anti-CD45 (clone 30-F11, BioLegend), anti-CD4 (clone RM4-5, eBioscience), anti-CD8 (clone 53-6.7, BioLegend), anti-NK1.1 (clone PK136, BioLegend), anti–granzyme B (clone QA16A02, BioLegend), and anti–IFN-γ (clone XMG1.2, BioLegend). For granzyme B and IFN-γ staining, cells were incubated in culture medium containing a cell activation cocktail (catalog 423303, BioLegend) at 37°C for 5 hours and then washed with permeabilization wash buffer (catalog 421002, BioLegend) before being stained with fluorescently labeled antibodies. Data collection and analysis were performed on a flow cytometer (Becton Dickinson).

C57BL/6 mice bearing either single or bilateral MC38 tumors received three fractions of 8 Gy radiation to the right hind flank tumors or were non-irradiated. From days 8 to 14, mice were intratumorally administered PBS, anti-ICAM-1 antibody (50 µg), or IgG isotype-matched control (50 µg) every other day. On day 17, the mice were euthanized, and the tumors and spleens were collected.
In a separate study, BALB/c mice with either single or bilateral 4T1 tumors received three fractions of 8 Gy radiation to the right hind flank tumors or were non-irradiated. From days 10 to 14, they received i.p. injections of PBS control or 200 μg of VS-6063 (Selleckchem) every other day. On day 16, the mice were euthanized, and the tumors were harvested.
Tumor samples were mechanically minced and digested in RPMI-1640 medium (Invitrogen) supplemented with 10 U/mL collagenase I, 400 U/mL collagenase IV, and 30 U/mL DNase (Yuanye Bio-Technology, Shanghai, China) at 37°C for 1 h. The spleens were dissected and filtered through a strainer. Red blood cells in the digested tumor samples and spleens were lysed using erythrocyte lysis buffer (BioLegend). Tumor-infiltrating cells and spleen cells were resuspended in PBS containing 0.5% w/v bovine serum albumin (BSA). The collected cells were then stained and analyzed using flow cytometry.