N n dimethylformamide (dmf)

Each mouse was treated by oral gavage administration of 0.2 ml of DMF (60 mg/kg; Selleck Chemicals; USA) [39 (link)–41 (link)] or 0.2 ml of vehicle control (10% DMSO + 10%Tween 80 + 40% PEG-300 + 50%PBS) once daily for 2 weeks following immunization (start DMF administration shortly after immunization on day 0). In this treatment method, DMSO was given to mice at dosages ranging from 0.88 g/kg to 1.22 g/kg, which was lower than the reported dosage that induced toxic effects such as liver degeneration, nephritis, and death in mice [42 (link), 43 (link)]. Another group of EAU mice were administered with DMF for 5 days (start DMF administration on day 10 after immunization) to test the therapeutic effect of DMF on established EAU. A Brief overview of the experimental design, grouping of mice, and timing of modeling and drug administration were shown (Additional file 1: Fig. S1A). For in vitro experiments, CDLN cells were isolated and administered with IRBP_1–20 at a concentration of 20 µg/mL, either individually or in combination with DMF at a concentration of 20 µM [28 (link), 44 (link)] for 72 h at 37 °C.

Cisplatin was purchased from Santa Cruz^® (reference sc-200896, diluted in saline), DMF was purchased from Selleckchem^® (reference S2586, diluted in 5% dimethyl sulfoxide [DMSO] + 30% Polyethylene glycol [PEG]400 + 5% Tween 80 + H₂O), Erlotinib was purchased from Selleckchem^® (reference S7786, diluted in 5% DMSO + 30% PEG400 + 5% Tween 80 + H₂O), and Fingolimod was purchased from Selleckchem^® (reference S5002, diluted in 5% DMSO + 30% PEG400 + 5% Tween 80 + H₂O).
Once the tumors reached an approximate volume of 100 mm³, the mice were randomized based on their tumor volume and body weight. Mice were treated with Cisplatin at 1 mg/kg via intraperitoneal (i.p.) route [84 (link),85 (link)], Erlotinib at 50 mg/kg via oral route [86 (link),87 (link)] Fingolimod at 5 mg/kg via oral route [88 (link),89 (link)], and DMF at 30 mg/kg via oral route [41 (link),54 (link)]. Drugs or vehicles were administrated five times per week until the end of the experiment.