Il 1β

All animal experiments were approved by the Institutional Animal Care and Use Committee at the National Cancer Center and carried out in accordance with the relevant guidelines and regulations. Ninj1 KO mice were provided by GT Oh^{20 (link),42 (link)}. C57BL/6J WT and Ninj1 KO mice were maintained under 12 h light/dark cycles at 22 °C and 60% humidity, and provided with food and water ad libitum. Eight-wk-old WT and Ninj1 KO mice were used in this study. To induce pulmonary fibrosis, 1 mg/kg of BLM (Carbosynth, Compton, UK) or PBS was intratracheally injected, and necropsy was performed at 1, 3, 5, 7, 14 and 21 days after injection. Before excision of lungs, cell-free bronchoalveolar lavage fluid (BALF) and BAL cells were collected at day 3, 7 and 21. as described in the previous report^{43 (link)}. The lungs collected at day 21 were fixed in 10% neutral formalin, and a paraffin block was generated. On days 1, 3, 5, 7, and 14, the right lungs were frozen in liquid nitrogen for RNA and protein analysis, whereas the left lungs were fixed in 10% neutral formalin for histological analysis. To measure levels of IL-1β and TGF-β1, enzyme-linked immunosorbent assay (ELISA kits; IL-1β, KOMA Biotech, Seoul, Korea; TGF-β1, AbFrontier, Seoul, Korea) was conducted using frozen lung tissue and cell-free BALF collected at day 7, following manufacturer’s instruction.

The cells (1 × 10⁵ cells/well, n = 3 per group) on a 96-well plate (Falcon) were pretreated with 500 μM of TUDCA for 1 h. In LPS containing TUDCA group, the cells were stimulated with LPS (1 μg/mL) containing 500 μM of TUDCA for 24 h. PGE₂ (R&D Systems, Minneapolis, USA), COX-2, iNOS (CUSABIO, Hubei, China), TNF-α, and IL-1β (Koma Biotech, Seoul, South Korea) were measured according to the each manufacturer’s directions.