Fig 2: DSC inhibits NF-?B activation. Colitis was induced as described in Materials and Methods and treated with or without DSC (50 mg/kg). A, Representative bands and quantitative analysis of p-NF-?B p65 in colonic tissues, NF-?B p65 was used as loading control. B, Colitis was induced as described in Materials and Methods and treated with or without lentiviral Nox4 shRNA. Representative bands and quantitative analysis of p-NF-?B p65 in colonic tissues, NF-?B p65 was used as loading control. BMDM was stimulated with or without DSC (50 µmol/L) as described in Materials and Methods. C, Representative bands and quantitative analysis of p-NF-?B p65 in BMDM and NF-?B p65 was used as loading control. D, Quantitative analysis of NF-?B p65 DNA binding activity in BMDM. Data shown are means ± SEM of n = 8 in each group. *P < 0.05 vs Control cell or mice (CTL), #P < 0.05 vs DSS-treated mice or LPS-stimulated cells

Fig 3: DSC inhibits Nox4-mediated redox imbalance in BMDM. BMDM was treated with or without DSC (50 µmol/L) or Nox4 siRNA as described in Materials and Methods. A, Representative bands and quantitative analysis of Nox4. B, Representative images and quantitative analysis of mitochondrial ROS production. C, Quantitative analysis of intracellular H2O2 generation. D, Quantitative analysis of NADPH oxidase activity. Data shown are means ± SEM of n = 8 in each group. *P < 0.05 vs Control cell (CTL), #P < 0.05 vs LPS-stimulated BMDM

Fig 4: DSC inhibits the activation of NLRP3 inflammasome in vivo and in vitro. A, Colitis was induced as described in Materials and Methods and treated with or without DSC (50 mg/kg). Representative bands and quantitative analyses of NLRP3, caspase-1 p20, cleaved IL-1ß and cleaved IL-18 in colonic tissues. The ß-actin was used as loading control (n = 8 in each group). B, Colitis was induced as described in Materials and Methods and treated with or without lentiviral Nox4 shRNA. Representative bands and quantitative analyses of NLRP3, caspase-1 p20, cleaved IL-1ß and cleaved IL-18 in colonic tissues. The ß-actin was used as loading control (n = 8 in each group). C, NLRP3 inflammasome was induced in BMDM as described in Materials and Methods and treated with DSC (50 µmol/L). Representative bands of NLRP3, ASC and caspase-1 in BMDM and caspase-1 p20 in supernatant (SN) by Western blot and quantitative analyses of IL-1ß in supernatant by ELISA. The ß-actin was used as loading control. Data shown are means ± SEM of n = 8 in each group. *P < 0.05 vs Control cell or mice (CTL), #P < 0.05 vs DSS-treated mice or LPS + nigericin-stimulated cells

Fig 5: DSC restores redox balance in colonic tissues. A, Colitis was induced as described in Materials and Methods and treated with or without DSC (50 mg/kg). Representative bands and densitometry analysis of Nrf2 nuclear translocation and HO-1 expression in colonic tissues. Histone H3 and ß-actin were used as loading control, respectively. B, Colitis was induced as described in Materials and Methods and treated with lentiviral Nox4 shRNA or lentiviral scrambled shRNA. Representative bands and densitometry analysis of Nrf2 nuclear translocation and HO-1 expression in colonic tissues. Histone H3 and ß-actin were used as loading control, respectively. BMDM was stimulated with or without DSC (50 µmol/L) as described in Materials and Methods. C, Representative bands and densitometry analysis of Nox4. ß-actin was used as loading control. D, Representative bands and densitometry analysis of Nrf2 nuclear translocation and cytoplasmic HO-1 expression in BMDM. ß-actin or histone H3 was used as loading control. Data shown are means ± SEM of n = 8 in each group. *P < 0.05 vs Control cell or mice (CTL), #P < 0.05 vs DSS-treated mice or LPS-stimulated cells