Efficacy validation on IBD mouse models for CsA (cyclosporin A). C57BL/6 mice were provided drinking water containing DSS for 7 consecutive days, and body weight changes were recorded throughout and scored clinically (A-D) at study endpoint, the colon weight and colon length were recorded. Two way-ANOVA (A-D) or one way ANOVA (E-F) followed by multiple comparison. Values are expressed as mean ± SEM. ****p<0.0001, ***p<0.001 , **p<0.01 , *p<0.05.

Histologic assessment of DSS-induced colitis in IBD mouse models. Photomicrographs of histopathological score and H&E staining cross-sections (10x and 40x magnification). Values are expressed as mean ± SEM. ****p<0.0001, ***p<0.001 , **p<0.01 , *p<0.05

Effects of anti-IL12/23p40 on DSS-induced acute colitis.
C57BL/6 mice received 3% DSS in drinking water for 5 days, CsA was injected to mice every day from day0 to day7, anti-IL12/23P40 was administered to mice on day2, day 6 and day 8 (A). Body weight and excreta were recorded every day(B, C). At the end, colon was collected for H&E staining, colon length (D) and weight (E) were recorded. CsA and anti-IL12/23p40 alleviated 3% DSS induced ulcerative colitis in this experiment. Two way-ANOVA followed by multiple comparison. Values are expressed as mean ± SEM. ****p<0.0001, ***p<0.001 , **p<0.01 , *p<0.05.

Efficacy validation on IBD(B-hCXCR2 mice) mouse models
C57BL/6 mice and B-hCXCR2 mice were provided drinking water containing DSS for 7 consecutive days, after which normal drinking water resumed. Body weight changes and clinical scores ( Weight loss score, stool hardness score, blood in stool score as well as total DAI score) were recorded throughout. Two way-ANOVA followed by multiple comparison, all group compared with G3. Values are expressed as mean ± SEM. ****p<0.0001, ***p<0.001 , **p<0.01 , *p<0.05.

Gross sampling and colon evaluation and histologic assessment of the colon of IBD mouse model. C57BL/6 mice and B-hCXCR2 mice were provided drinking water containing DSS for 7 consecutive days, after which normal drinking water resumed. At study endpoint, colon length (A) and colon weight (B) were measured. Colon tissue were used for H&E staining (C). One-way ANOVA followed by multiple comparison, all group compared with G3. Values are expressed as mean ± SEM.

C57BL/6 mice and B-hTL1A/IL23A/IL12B mice were provided drinking water containing different concentration of DSS for 7 consecutive days. Body weight changes and clinical scores (Weight loss score, stool hardness score, blood in stool score as well as total DAI score) were recorded throughout. Two way-ANOVA followed by multiple comparison, all group compared with G1. Values are expressed as mean ± SEM. ****p<0.0001, ***p<0.001 , **p<0.01 , *p<0.05.

C57BL/6 mice and TL1A/IL23A/IL12B humanized mice (B-hTL1A/IL23A/IL12B mice) were provided drinking water containing different concentration of DSS for 7 consecutive days. At study endpoint, colon length and colon weight were measured. And colon tissue were used for H&E staining. One way ANOVA followed by multiple comparison compared to G1. Values are expressed as mean ± SEM. ****p<0.0001, ***p<0.001 , **p<0.01 , *p<0.05.

DSS induced chronic colitis. Mice were repeatedly exposed to 4 cycles of DSS. Each cycle contained 2% DSS for 5 days, followed by water for the next 2 days (A). Body weight, colon weight and colon length were recorded (B). Histopathological staining by H&E (C) and Sirius Red (D). Colon was collected for histology analysis. Mice received DSS showed lower body weight, higher colon weight and shorter colon length than mice received water. Multiple T-test. Mean ±SEM, n=5, ****p<0.0001, ***p<0.001 , **p<0.01 , *p<0.05.

• Experimental Animals: C57BL/6N, 7-10 weeks old, female
• Detection Indicator: Body Weight, Colon Weight, Colon length
• Modeling reagent: TNBS (trinitrobenzene sulfonate)

Effects of CD4-GK1.5-mlgG2a on TNBS-induced acute intestinal inflammation. Animals were presensitized with TNBS solution on shaved skin area on day 0, and then received TNBS intrarectal administration on day 7. Body weight (A) and body weight change (B) were recoded every day, CD4-GK1.5-mlgG2a (3, 10 mg/kg, i.p.) were administered on day 6. Mice were sacrificed on day 10 and colon length (C) and weight (D) were recorded. Two-way ANOVA or One-way ANOVE followed with Dunnett multiple comparisons, all groups compared with each other. Values are expressed as mean ± SEM. ****p<0.0001, ***p<0.001, **p<0.01 , *p<0.05.

Efficacy of PF06480605 on TNBS-induced acute IBD on B-hTL1A mice. Animals received TNBS intrarectal administration on day 0. For TA administration, 10 of 25 mg/Kg PF06480605 were administered on day -1, day 2 and day 5. Schematic diagram of the scheme (A). Survival rates (B) were calculated and body weight (C-D) were recorded every day. Two-way ANOVA followed with Dunnett multiple comparisons, all groups compared G2. Values expressed as mean ± SEM. ****p<0.0001, ***p<0.001, **p<0.01 , *p<0.05.

Animals received TNBS intrarectal administration on day 0. Body weight were recorded every day and survival rates were calculated. For TA administration, 10 of 25 mg/Kg PF06480605 were administered on day -1, day 2 and day 5. On day 7, mice were sacrificed and colon weight (A) and length (B) were recorded. Colon tissue was later used for H&E staining. One-way ANOVA followed with multiple comparisons. All groups compared with G2. Values expressed as mean ± SEM. ****p<0.0001, ***p<0.001, **p<0.01 , *p<0.05.

Efficacy of T cell transfer inducing IBD models.
The spleens of Balb/c mice were used to obtain CD4+CD25- T cells or CD4+ T cells using commercial kits. Animals in G2 received 3×105 CD4+ T cells injection while animals G3-G5 received same quantity of CD4+CD25- T cells. For treatment, animals in G4 received weekly administration of 10 mg/kg anti-TNFα and animal in G5 received weekly administration of 10 mg/kg anti-CD4. Two-way ANOVA or One-way ANOVA followed with multiple comparison, Every group compared with G3. Values expressed as mean±SEM. ****p<0.0001, ***p<0.001, **p<0.01, *p<0.05.