Cell experiment:

Luciferase assays are carried out using the H1L7.5c3 cells. At the indicated times (0.5, 2, 4, 6, 12, 18, 24 h) and concentrations (0.1, 1, 10, 100 μM) of exposures to Kynurenine, cells are removed from incubation and allowed to equilibrate to room temperature for 15min. After equilibration, the medium is removed and the cells are washed twice with at room temperature with DPBS. The cells are lysed with 20 μL/well 1× Passive Lysis Buffer and shaken for 20min at room temperature. Luciferase activity is recorded using an Luminometer Microplate Reader[1].

Animal experiment:

Rats[3]The effects of increasing doses of L-kynurenine with or without probenecid on concentrations of kynurenic acid in cerebral cortex are examined in 7-day-old rats. Six animals are examined in each group. Animals are treated with L-kynurenine at doses of 100, 200, 300, and 400 mg/kg or kynurenine, 200 mg/kg with probenecid, 50 mg/kg. Animals are killed at 1 h, the brains promptly removed, and the cerebral cortex is dissected and placed in 0.5 mL of chilled 0. 1 M HCl. Kynurenic acid measurements are made by high-performance liquid chromatography with fluorescence detection. Protein measurements are made using a fluorometric assay[3].