Figure 5

Bleomycin-induced pulmonary fibrosis was attenuated by treatment with PI3K inhibitor. (A) Flow chart of the experimental procedure. Mice with intratracheal administration of bleomycin (2 U/kg) or saline (vehicle) were treated with or without PI3K inhibitor, LY294002 (LY, 50 mg/kg, i.p.) before (Upper: prevention) or 7 days (Lower: treatment) after bleomycin intratracheal instillation. The mice were sacrificed 14 days later and the lung specimens were harvested for histological analysis with (B) HE, (C) picro pirius red and (D) Masson’s trichrome staining followed by (C and D Right) quantification, (E) the results of total collagen assay, (F and G) immunohistochemical analysis, and (H,I) Western blot analysis for the expression of proteins associated with (H) AKT and (I) ER stress after treatment of PI3K inhibitor, LY294002, in control, prevention and treatment groups. (Cropped blots are displayed; Full-length blots are presented in Supplementary Figure, labeled Figure S5).