A5486 25mg

Azoxymethane (AOM, 10mg/kg, Sigma-Aldrich- A5486-25MG) in saline was injected intraperitoneally (IP) 2 days in advance of three cycles of 3.5% dextran sodium sulfate (DSS, MP Biomedicals- 160110, molecular weight: 36,000–50,000). Each cycle consisted of 5 days of 3.5% DSS in the drinking water, followed by 16 days of regular water. Mice were euthanized at various time points (“baseline”; at the end of each DSS treatment, designated as “colitis”; one week after each DSS cycle, designated as “recovery”, with the end of the 3^rd cycle being designated as “cancer”). This AOM-DSS colitis model leads to predictable and reproducible colitis and cancer [15 ]. Disease activity index (DAI) was recorded, a colonoscopy was performed 1 week after each DSS treatment, and mice were euthanized at various time points (colitis, recovery, cancer) with colon tissue either stored in liquid nitrogen or placed in formalin. Disease activity index was calculated based on weight loss (1 = 0–10%, 2 = 10–15%, 3 = 15–20%, 4 = >20%), stool consistency (1 = solid, 2 = loose, 3 = wet, 4 = diarrhea), and hematochezia (1 = no blood, 2 = slight blood, 3 = blood, 4 = significant hematochezia).

The azoxymethane/dextran sodium sulfate (AOM/DSS) model was injected intraperitoneally with 12.5 mg/kg azoxymethane (AOM, 10 μL/g, A5486‐25MG, SIGMA, USA) solution on Day 1 of Week 1. After resting for 1 week, mice were fed drinking water containing 2.5% dextran sodium sulfate (DSS, MB5535, meilunbio, China) for 1 week, then switching to regular drinking water for 2 weeks and cycling three times, as shown in Figure 1A. In the AOM/DSS induced model, anti‐PD‐1 (BE0146, Bio X Cell, USA) was injected intraperitoneally on Days 50, 57, and 64, respectively, as shown in Figure 5A. The APC^min/+ DSS model was given drinking water containing 2.0% DSS to all animals for 1 week, followed by switching to regular drinking water for 2 weeks and cycling two times, as shown in Figure 2A. We divided tumors into three groups: small tumors, <1 mm; medium tumors, 1 mm ≤ and ≤2 mm; large tumors, >2 mm. Tumor load was calculated according to the following formula: tumor load = (number of small tumors) × 1 + (number of medium tumors) × 2 + (number of large tumors) × 3.¹⁶

For induction of CAC³² , 6-week-old male Rig-i^fs/fs and wild-type (WT) mice were used. 7 days after a single intraperitoneal injection with 10 mg/kg of Azoxymethane (AOM) (Sigma, A5486-25MG), mice were treated with 2% (weight/volume) dextran sulfate sodium (DSS) (MP, 0216011090) for 3 cycles. 1 cycle includes 7 days DSS treatment and followed 14 days resting (drinking regular water). Mice were sacrificed at day 90 before tumor size reaches the maximal tumor burden (10% of mouse weight or 2000 mm3 at the endpoint) permitted by ECPKUHSC