C57BL/6J mice of either sex (8–12 weeks old) were weighed 2 days prior to the procedure; weight and stool content/consistency were then monitored daily throughout the treatment. Drinking water supplemented with 3% DSS (40,000 MW, Alfa Aesar) was administered with control mice receiving the same drinking water without DSS. The mice received DSS‐treated water for 5 days after which it was replaced with normal drinking water for a further 2 days. The experimenter was blinded, and solutions labeled A and B being unblinded after results were fully analyzed. Oral administration of DSS leads to weight loss, diarrhea, and blood in the stool, which was scored to produce a disease activity index (DAI) (Table 3 ) (Manicassamy & Manoharan, 2014 ). On day 7, mice were killed, and relevant tissue samples, Wallace macroscopic score (Table 4 ; Fang et al., 2002 ), and measurements were obtained. An approximate 1‐cm section of colon was removed about 4 cm from the anus and fixed in 4% paraformaldehyde for 4 hr and cryoprotected in 30% sucrose overnight. The tissue was then embedded in O.C.T mounting media (VWR Q‐path Chemicals), snap frozen in liquid nitrogen, and stored at −80°C until needed.
Dextran sulfate sodium (dss)
Manufactured by Thermo Fisher Scientific
Sourced in United States, Germany, United Kingdom
The DSS is a laboratory equipment product designed for sample preparation and handling. It provides a consistent and controlled approach to sample processing, ensuring reproducible results. The core function of the DSS is to assist in the preparation and manipulation of samples for further analysis or testing.
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Lab products found in correlation
Formic acid, by Thermo Fisher Scientific (6 mentions)
Acetonitrile, by Thermo Fisher Scientific (4 mentions)
Rnalater, by Thermo Fisher Scientific (4 mentions)
Sodium chloride (nacl), by Merck Group (3 mentions)
Veterinary endoscopy, by Storz (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Dextran sulfate sodium (dss), by Merck Group (2 mentions)
Dynabeads protein g immunoprecipitation kit, by Thermo Fisher Scientific (2 mentions)
Dextran sulfate sodium (dss), by MP Biomedicals (2 mentions)
Mg132, by Merck Group (2 mentions)
Criterion precast midi protein gel, by Bio-Rad (2 mentions)
Pkm2 antibody, by Santa Cruz Biotechnology (2 mentions)
Ag 25b 0006 c100, by Adipogen (2 mentions)
Protease inhibitor cocktail, by Merck Group (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Methylamine, by Merck Group (2 mentions)
Urea, by Merck Group (2 mentions)
Cacl2, by Merck Group (2 mentions)
Trypsin, by Promega (2 mentions)
Asp n, by Promega (2 mentions)
152 protocols using dextran sulfate sodium (dss)
1
Dextran Sulfate Sodium-Induced Colitis Model in Mice
2
Acute and Chronic Colitis Models in Mice
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8-week-old mice were used in this investigation. For the acute TNBS (Sigma-Aldrich)-induced colitis model, 1% TNBS solution was applied to the back skin of mouse for 8 days for pre-sensitization. Overnight-fasted presensitized mice were treated with 100 mg/kg TNBS dissolved in 50% alcohol under anesthesia via intrarectal injection for 3 days, control mice were subjected to 50% alcohol. For the DSS-induced colitis model, mice were treated with 7 days of 2.5% DSS (Fisher Scientific) water and 2 days of tap water.
3
Culturing and Treating Cell Lines
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Drosophila S2 cells (Invitrogen, Carlsbad, CA) were maintained in Schneider media (Invitrogen) with 10% fetal bovine serum (FBS), human U937 cells were grown in RPMI1640 with 10% FBS, and primary human lung fibroblasts were grown in DMEM with 10% FBS. Mouse embryonic fibroblasts and splenic monocytes were isolated from flotillin-1−/− mice (Ludwig et al., 2010 (link)). For studies using chloroquine or MG132, U937 cells were grown on 100-mm dishes and treated with 50 μM or different concentrations of chloroquine (Sigma-Aldrich, St. Louis, MO) as indicated in Figure 4 for 12 h or 20 μM of MG132 (Sigma-Aldrich) for 24 h before immunoblot analysis. For studies using DSS (Thermo Scientific, Waltham, MA), U937 cells were grown on six-well plates and treated with DSS for different periods as indicated in Figure 4E .
4
DSS-Induced Colitis in Mice
5
Colitis Induction in Murine Models
6
Antioxidant and Cytotoxicity Assessment
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The following substances were used: 2,2-diphenyl-1-picrylhydrazyl, 5,5′-dithiobis (2-nitrobenzoic acid), bovine serum albumin, glutathione, butylated hydroxytoluene, Griess reagent, MTT, pyrogallol, xylenol orange (all from Sigma, St. Louis, USA), absolute ethanol, acetic acid, ascorbic acid, ferrous ammonium sulfate, hydrochloric acid, formaldehyde, hydrogen peroxide, methanol, sodium acetate, trichloroacetic acid (Vetec, Rio de Janeiro, RJ, Brazil), sulfuric acid, dimethyl sulfoxide and N,N-dimethylformamide (DMSO, Synth, Diadema, SP, Brazil), Dulbecco's Modified Eagle Medium (DMEM, Vitrocell, Campinas, SP, Brazil), fetal bovine serum (FBS, Gibco), and dextran sulfate sodium (Alfa Aesar, Heysham, Lancashire, UK).
7
Colon Cancer Induction in Mice
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Mice were treated with 10 mg/kg/animal of azoxymethane (AOM; Sigma) via intraperitoneal injection, followed by oral inoculation with 1 × 109 CFU of E. coli strains at 24 h postinjection. Supplementation with 3% dextran sulfate sodium (DSS; Alfa Aesar) in the drinking water was continuously provided for a week postinfection. Two subsequent weeklong DSS supplementations were given in between 3-week rests during the 9-week experiment. The AOM/DSS short-term treatment samples were collected at day 8 of the first DSS treatment.
8
Inflammatory Bowel Disease Cytokine Assay
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Dextran sulfate sodium (MW ca 40,000 Da, cat no. J63606) was purchased from Alfa Aesar. RIPA Lysis Buffer (cat no. 89900) and Halt™ Protease Inhibitor Cocktail (cat no. 87786) were purchased from Thermo Scientific™. Dopamine hydrochloride (cat no. H8502) and Phorbol 12-myristate 13-acetate (PMA, cat no. 8139) were purchased from Sigma Aldrich. DNase-I (cat no. 10104159001), Collagenase D (cat no. COLLD-RO), and 1,4-dithiothreitol (DTT, cat no. DTT-RO) were purchased from Roche. Mesalamine (cat no. PHR1060) was purchased from Supleco. TRIzol™ Reagent (cat no. 15596026), RNAlater™ Stabilization Solution (cat no. AM7020), DNase I Buffer (cat no. AM8170G), Ambion™ DNase I (cat no. AM2222), and UltraPure™ Salmon Sperm DNA Solution (cat no. 15632011) were purchased from Invitrogen™. Percoll™ (Density 1.130 g/mL, cat no. 17089101) was purchased from Cytiva. Poly(lactic-co-glycolic acid) (50/50 ratio, inherent viscosity 0.55–0.75, acid terminated, cat no. B6013-2P) was purchased from LACTEL® Absorbable Polymers. 4arm polyethylene glycol (HCL salt, MW 5000, cat no. 4ARM-NH2 5000) was purchased from JenKem Technology.
9
Azoxymethane and Dextran Sulfate Sodium Induced Colorectal Cancer in Mice
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Mice received an intraperitoneal (i.p.) injection of 12.5 mg/kg azoxymethane (AOM) (Sigma, St. Louis, MO, USA). Five days later, 2% dextran sulfate sodium (DSS, MW:40,000, Alfa Aesar, Tewksbury, MA, USA) in drinking water was administered ad libitum for 7 days. Mice were then provided regular water for 14 days and subjected to two more DSS cycles. To examine the early and late transformative steps in CAC, the mice were killed on days 20, 40 (early tumor development), and 77 (late tumor development) after AOM injection. Throughout the experiment, Disease Activity Index (DAI) scores were given to evaluate the clinical progression of CAC. DAI scores were calculated as the sum of changes in weight loss, compared to initial weight, stool consistency, and bleeding. During the necropsy, the colons were removed, weighed, and submitted for macroscopic inspection and histopathological examination.
10
Azoxymethane-Induced Colorectal Cancer Model
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Azoxymethane was purchased from Sigma-Aldrich chemical company, St. Louis, USA. Dextran sulfate sodium (MW ca 40,000) was purchased from Alfa Aesar, UK. All other chemicals and reagents used were of analytical grade. ZJ was purchased from Gong-guan Farmer's Cooperative, Miaoli, Taiwan.
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