TUG1 Modulation in Lung Cell Models

BEAS-2B (human lung epithelial cells) and HFL1 (human lung fibroblasts) cells were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA). BEAS-2B was cultured in LHC-8 medium without gentamicin (Thermo Fisher Scientific, Waltham, MA, USA) and HFL1 was cultured in F-12K medium (Thermo Fisher Scientific). Penicillin (100 U/mL), streptomycin (100 μg/mL), and 10% fetal bovine serum (FBS; Thermo Fisher Scientific) were added to all the culture media. Cell lines were cultured with 5% CO₂ at 37°C. For cell treatments, we used 2 ng/mL TGF-β (R&D Systems, Inc., Minneapolis, MN, USA) for 48 h. Transient transfection was assayed using Lipofectamine 3000 (Thermo Fisher Scientific) according to the manufacturer’s protocol. NC (negative control) siRNA and TUG1-siRNA were synthesized. The siRNA sequences used to target TUG1 were 5′-GCU UGG CUU CUA UUC UGA AUC CUU U-3′ (sense) and 5′-AAA GGA UUC AGA AUA GAA GCC AAG C-3′ (antisense).29 (link) BEAS-2B and HFL1 cells were transfected with Lipofectamine 3000 (Thermo Fisher Scientific). Cells were cultured for 48 h after transfection and then were analyzed by cell viability assay and Western blot analysis.