Fig 2: TRIM67 deletion represents the overexpression of pro-inflammatory cytokines in ileum and colon. (A,B) Relative mRNA expression of TNF-a (n = 5), IL-6 (n = 5), IL-1ß (n = 4) and IL-2 (n = 4) in ileum and colon. (C,D) Western blots and quantification to check protein level of TNF-a, IL-6, and IL-1ß in ileum and colon (n = 3). (E–G) Relative mRNA expression of TNF-a, IL-6, and IL-1ß in Myc-TRIM67 and Myc cell lines supplemented with NaHCO3 and PA, “-”: no addition, “ + ”: addition (n = 4). (H) Western blots and quantification display the protein level of TNF-a, IL-6, and IL-1ß in Myc and Myc-TRIM67 groups supplemented with NaHCO3 and PA, “-”: no addition, “+”: addition (n = 3). The data are expressed as mean ± SEM. Significance was determined by one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001).

Fig 3: TRIM67 knockout mice represent suppressed antioxidant capacity in ileum and colon. (A) The mRNA expression of SOD-1 (n = 4) and SOD-2 (n = 4) in ileum. (B) Relative mRNA expression of CAT (n = 4), SOD-1 (n = 4) and SOD-2 (n = 5) in colon. (C,D) SOD activity and GSH level in ileum and colon to check the antioxidant activity (n = 8). The data are expressed as mean ± SEM. Significance was determined by one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001).

Fig 4: TRIM67 deletion has no effect on body weight and exacerbates the pathological damage. (A) Average weekly feed intake of mice during modeling WT CTR compared to WT HFD (n = 8), * p < 0.05, ** p < 0.01, *** p < 0.001; KO CTR compared to KO HFD, # p < 0.05, ## p < 0.01, ### p < 0.001; WT CTR compared to KO CTR, & p < 0.05, && p < 0.01, &&& p < 0.001. (B) Average weekly weight gain of mice WT CTR compared to WT HFD (n = 11), * p < 0.05, ** p < 0.01, *** p < 0.001; KO CTR compared to KO HFD, # p < 0.05, ## p < 0.01, ### p < 0.001. (C) H&E results of ileum indicating the pathological damage in different groups as compared to WT CTR diet group. (D) Colon H&E staining results displaying the difference in lymphocyte number, height of intestinal mucosa and colon diameter in different groups. (E,F) Quantification of villus height and crypt depth in ileum (n = 5). (G–I) The quantification of lymphocyte number, height of intestinal mucosa and diameter in colon (n = 5). (J,K) PAS staining results in ileum and colon and graphical representation of goblet cell count. Black arrows showing the goblet cells in ileum and colon (n = 5). The data are expressed as mean ± SEM. Significance was determined by one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001).

Fig 5: TRIM67 knockout mice display the lower expression of anti-inflammatory cytokines and HFD feeding exacerbates the condition. (A,B) Relative mRNA expression of anti-inflammatory cytokines IL-4 (n = 4) and IL-10 (n = 5) in ileum and colon. (C,D) Western blots and quantification show protein level of IL-4 and IL-10 in ileum and colon (n = 3). (E) Relative mRNA expression show expression of IL-10 in Myc-TRIM67 and Myc group supplemented with NaHCO3 and PA, “-”: no addition, “+”: addition (n = 4) (F) Western blots and quantification of IL-10 and IL-4 proteins in cell lines, “-”: no addition, “+”: addition (n = 3). The data are expressed as mean ± SEM. Significance was determined by one-way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001).