Omni th 115 homogenizer

Lung adenocarcinoma and normal lung tissue samples, obtained from resected lung cancer specimens from patients who had surgery at MD Anderson Cancer Center, were homogenized using Omni plastic disposable probes and an Omni (TH‐115) homogenizer (Omni International, Warrenton, VA) for 1 minute on dry ice. Total RNA was isolated from the cells using the Direct‐zol™ RNA Miniprep kit from Genesee Scientific (San Diego, CA) and samples homogenized using Trizol reagent according to the manufacturer's instructions. Total RNA was quantified using the Nanodrop 1000 spectrophotometer (Thermo Scientific, Wilmington, DE). RNA quality was assessed based on RNA integrity numbers generated by the Agilent Bioanalyzer 2000 (Agilent Technologies, Palo Alto, CA) according to the manufacturer's instructions.

Human NSCLC cell line A549 was obtained from the American Type Culture Collection and maintained in RPMI 1640 supplemented with 10% fetal bovine serum, penicillin (100 units/mL), and streptomycin (100 Ag/mL; Invitrogen, Corp.) at 37°C in a humidified atmosphere with 5% CO₂. Previous studies showed that α5-nAChR is expressed in several NSCLC cell lines (Lam et al., 2007 (link)). Total RNA was isolated from cells using the RNeasy kit from Qiagen according to the manufacturer’s instructions. Lung adenocarcinoma and normal lung tissue samples were homogenized using Omni plastic disposable probes and an Omni (TH-115) homogenizer (Omni International) for 1 min on dry ice after which total RNA was isolated using Trizol reagent according to the manufacturer’s instructions. Total RNA was quantified using a Nanodrop 1000 spectrophotometer.