Lung specimens
cancer
CANCER, LUNG
Lung specimen from 14 different individuals was obtained by postsurgical resection of anonymous lung tissue (typically for cancer resections). The lung sections from lung regions having normal macro- and microscopic appearance were used. Several small pieces were cut from all samples and 5–6 pieces/well/condition were cultured for 4 days in 24 well plates containing 1 ml of RPMI medium (supplemented with 10% fetal calf serum, 4 mM L-glutamine, 100 U/ml penicillin, and 100 μg/ml streptomycin), IFN β (1000 UI/ml FP1201; Faron Pharmaceuticals), or placebo with and without hydrocortisone (40 µg/ml, Solucortef, Pfizer). After culturing, all pieces in each well were collected and frozen in OCT. All 14 samples were used for staining IFNAR1/IFNAR2 and 5 CT and 5 TT samples for STAT1, pSTAT1, STAT2, pSTAT2, and CD73 stainings.DNA was extracted from 3 to 5 frozen lung tissue sections (20 µm) with NucleoSpin DNA Rapid Lyse Kit (Macherey–Nagel). TaqMan SNP genotyping assays C_2443264_10 and C_11354003_30 (Thermo Fisher Scientific) for rs9984273 and rs2236757, respectively, were used in genotyping. The reactions were performed as in TaqMan SNP genotyping manuals (ThermoFisher). The plate was run on a QuantStudio 3 Real-Time PCR machine (ThermoFisher) with a run protocol for SNP genotyping as per manual. The results were analyzed using the Applied Biosytems® analysis modules for genotyping in Thermo Fisher Cloud computing platform (ThermoFisher Scientific).