Recombinant mouse il 1β

Nobiletin (purity ≥ 98%), collagenase-II, and dimethyl sulfoxide (DMSO) were obtained from Solarbio (Beijing, China). Recombinant mouse IL-1β was purchased from Novoprotein (China). Primary antibodies directed against A disintegrin and metalloproteinase with thrombospondin motifs-5 (ADAMTS5), collagen II, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were purchased from Abcam (Cambridge, MA, USA), while primary antibodies against iNOS, MMP-3, MMP-13, cyclooxygenase-2 (COX-2), p-IkBα, IkBα, p-p65, and p65 were obtained from ProteinTech (Wuhan, China). Cell-Counting Kit-8 (CCK-8) was purchased from Dojindo (Kumamoto, Japan). Fetal bovine serum (FBS), Dulbecco’s modified Eagle’s medium (DMEM)/F12, 0.25% trypsin ethylenediaminetetraacetic acid (trypsin–EDTA), and bovine serum albumin (BSA) were purchased from Healthcare Life Sciences (Hyclone; Logan, UT, USA). Griess reagent and Bicinchoninic acid radioimmunoprecipitation assay lysis buffer were purchased from Solarbio (Beijing, China). The NF-kB inhibitor pyrrolidine dithiocarbamate (PDTC) was obtained from Abcam (Cambridge, UK). Goat anti-mouse horseradish peroxidase conjugates and goat anti-rabbit were purchased from Jackson ImmunoResearch (West Grove, PA, USA). Griess reagent and PGE2 ELISA kits were obtained from Bio-Swamp Life Science (Shanghai, China).

Recombinant mouse IL-1β (Novoprotein, Shanghai, China) or PBS as negative control was injected into Gsdmd^-/- mice intraperitoneally at a concentration of 0.5 μg or 1 μg per mouse in 200 µl sterile PBS on Days 1, 2, 3, 4, 5, and 6 during DSS-induced colitis.
Recombinant mouse IL-18 (Novoprotein) or PBS as negative control was injected into Gsdmd^-/- mice intraperitoneally at a concentration of 1 μg per mouse in 200 µl sterile PBS from Day 1 to Day 8 during DSS-induced colitis.