Mouse Model Validation of Drug Efficacy

Acute colitis was induced in mice by 3% DSS in drinking water ad libitum for seven days and followed with four days of normal distilled water. Control mice were fed with distilled water for 11 days. The colitis group mice were randomly grouped and intraperitoneally injected with 100 µL vehicle (0.01% DMSO) or BFA (2.5, 5, and 10 mg/kg) at day 4 and 6 post-DSS. Mice were evaluated daily for changes of body weight. At day 9 and 11, mice were euthanized to harvest colons and blood for further analysis. Colon lengths were measured at autopsy. Mouse colon biopsies were lysed in HTAB buffer to determine the MPO activity and in RIPA buffer for IL-1β secretion (ELISA) and cGAS expression (immunoblotting). Serum was separated from clotted blood by centrifugation. Serum AST and ALT were measured by ELISA (IT5530 and IT5508, G-Biosciences) according to the manufacturer's instructions.