Bleomycin‐induced pulmonary fibrosis model mice

Sex‐, age‐, and weight‐matched C57BL/6J mice (8–10 weeks of age) were anesthetized using 13 μL·g⁻¹ of 4% tribromoethanol. A maximal 1‐cm midline cervical incision was made to expose the trachea, followed by intratracheal instillation of PBS as a vehicle or 1.2 U·kg⁻¹ of bleomycin (Nippon Kayaku, Tokyo, Japan). The cervical incision was closed with n‐butyl cyanoacrylate (Vetbond, 3M Health Care, St. Paul, MN, USA), and the mice were returned to their cages to recover. Mice were sacrificed 7 days after instillation to obtain BALF and serum samples. All experimental procedures were performed according to guidelines of the Committee on Ethical Use of Laboratory Animals of the Osaka University Medical Department.