Female c57bl 6 mice

Wild-type (WT) female C57BL/6 mice (eight-week-old) in the study were obtained from SPF Biotechnology Co., Ltd of Beijing, China. Female Nlrp3^−/− mice were supported by Dr. Tao Li from National Center of Biomedical Analysis (Beijing, China). All mice were maintained under a pathogen-free condition (22 ± 2 °C) and held under a 12-h dark/light cycle. We tried our best to minimize the suffering as well as the number of animals used. When assessing experimental outcomes, the investigators were blinded to the treatments.

Female C57BL/6 mice (6-8 weeks) were purchased from SPF Biotechnology Co., Ltd. (Beijing) (Stock number: B204). Female BALB/c mice (6–8 weeks) were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd (Stock number: 211). Mice were housed under conditions of a light/dark cycle of 12 h, an ambient temperature of 25 ± 2 °C, and a humidity of 60 ± 10%. All the animals were treated according to the Guide for Care and all relevant animal experiments were performed in compliance with the animal management rules of the Ministry of Health, People’s Republic of China and Animal Experiment Ethics Review Board of Shandong University. When the tumour volumes reached ~2000 mm³, the mice were euthanized by rapidly dislocation of the head and neck.

Female C57BL/6 mice aged between six to eight weeks were purchased from SPF Biotechnology Co. Ltd. (Beijing, China). B16F1 cells were inoculated subcutaneously into C57BL/6 mice at a density of 5×10⁵. Nine days after inoculation, mice were divided into four groups (PBS group, doxorubicin group, gemcitabine group, and combined treatment group) and the corresponding groups were PBS (volume 100ul/mice), doxorubicin (5 mg/kg), gemcitabine (25 mg/kg), and a combination of doxorubicin and gemcitabine for six days, respectively. Doxorubicin (S1208) and gemcitabine (S1149) were purchased from Selleck Chemicals (Houston, TX). After initiating the treatment, tumor length and width were measured daily, and tumor volumes were quantified as (length×width×height)/2. All mice were observed, treated, and euthanized according to the protocols of the Animal Ethics and Welfare Committee of Tianjin Medical University Cancer Institute and Hospital. The animal study protocol was approved by the Animal Ethics and Welfare Committee of Tianjin Medical University Cancer Institute and Hospital (protocol code AE-2021030, September 9, 2021).