In model of pulmonary fibrosis mimosine was administed at doses 25 and 50 mg/kg. Pulmonary fibrosis was induced by tracheal instillation of 5 mg/kg bleomycin. Mimosine was first dissolved in Tris-buffer at pH 8.9, and then pH was adjusted to 7.2 with 1 N HCl. Before administration, this solution was freshly diluted by 0.9% physiologic saline to achieve the required concentration. L-Mimosine was administered via subcutaneous injection once daily from day 1 to day 28 after BLM or saline treatment.

The effect of mimosine on asynchronously proliferating HeLa cells was analyzed by flow cytometry. HeLa-S3 cells were cultured as proliferating monolayers on 145-mm plates in DMEM, supplemented with 10% FCS, 10 U/ml penicillin, and 0.1 mg/ml streptomycin. Mimosine stock solution was prepared at 10 mM in DMEM and filtered through 20-mm filters. Synchronization of HeLa cells by mimosine was achieved by adding mimosine from the stock solution directly to the cells at the concentrations specified. Cell synchronization was determined by flow cytometry of isolated nuclei. One million nuclei were directly stained with propidium iodide (5 mg/ml in PBS containing 0.4% Triton X-100) and analyzed by FACScan (Becton Dickinson) using the Lysis II-software.