Bronchial epithelial cell growth medium bullet kit begm

Cell culture-grade water, phosphate buffered saline (PBS), heat-inactivated fetal bovine serum (FBS), N-acetyl cysteine (NAC) and Bradford protein assay reagents were purchased from Sigma (St. Louis, MO). Bronchial epithelial cell growth medium bullet kit (BEGM) was obtained from Lonza (Walkersville, MD). Dulbecco's Modified Eagle Medium (DMEM), penicillin/streptomycin mix and trypsin-EDTA were from Invitrogen (San Diego, CA). OptEIA™ Human IL-6 and IL-8 ELISA kits and type I rat tail collagen were purchased from BD Bioscience (San Diego, CA). Trypan blue solution and chemiluminescent substrate were from HyClone (Waltham, MA) and Thermo Fisher Scientific (Rockford, IL), respectively. Antibodies against phospho-p38, total p38, COX-2, and cell lysis buffer were obtained from Cell Signaling Technology (Beverly, MA). Monoclonal anti-heme oxygenase-1 (HO-1) Ab and PGE2 ELISA kit were from Enzo Life Sciences (Farmingdale, NY). R,S-Sulforaphane (SFN) and SB203580 were purchased from LKT Laboratories (St. Paul, MN) and EMD Millipore (Bedford, MA), respectively. RNeasy Mini Kit was from Qiagen (Valencia, CA). High Capacity cDNA Reverse Transcription Kit was purchased from Applied Biosystem (Grand Island, NY).

Non-cancerous, immortalized human lung bronchial epithelial cell line BEAS-2B was purchased from Zhong Qiao Xin Zhou Biotechnology (Shanghai, China). Normal fetal lung fibroblast cell HFL1; human LUAD cell lines Calu-3, NCI-H1975, and NCI-H1395; LUSC line NCI-H520; and other NSCLC cell lines A549 and NCI-H1299 were purchased from Procell (Wuhan, China). LUSC lines NCI-H226 and SK-MES-1 were purchased from Shanghai Chinese Academy of Science Cell Bank (Shanghai, China). BEAS-2B was grown in Bronchial Epithelial Cell Growth Medium BulletKit (BEGM; Lonza, Basel, Switzerland). HFL1 was cultured in Ham’s F-12K medium (Gibco, Pittsburgh, PA, USA). Calu-3 and SK-MES-1 were cultured in Eagle’s minimum essential medium (MEM; Gibco). NCI-H1975, NCI-H1395, NCI-H1299, NCI-H520, and NCI-H226 were maintained in RPMI-1640 (Gibco). All cell lines, except for BEAS-2B, were supplemented with 10% fetal bovine serum (FBS), streptomycin (100 μg/mL), and penicillin G (100 U/mL; Gibco). All cell lines were authenticated by short tandem repeat (STR) profiling. Cells were incubated at 37°C in a humidified atmosphere with 5% CO₂.