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TNBS colitis involves skin sensitization of mice with the haptenating agent TNBS 1 week prior to single rectal administration to mice of TNBS in ethanol (46). When TNBS couples with proteins with high molecular weight, it presents them as immunogenic to the host immune system (47). Mice were sensitized by epicutaneous application of TNBS (1% in 100% ethanol, Sigma-Aldrich) or vehicle (100% ethanol) 1 week prior to rectal instillation of TNBS (5% in 40% ethanol) or vehicle (40% ethanol), as previously described (46). Weight changes in mice were monitored for 1 week, and tissue was collected and processed as described for DSS colitis.
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