Pck 26

All tissue samples were procured by the Human Tissue Procurement Service shared resource at the Winship Cancer Institute of Emory University in accordance with the approved institutional review board protocol. Tissues were digested for 3 hours in digestion buffer [DMEM/F-12 supplemented with 10 mM Hepes, 2% bovine serum albumin, 1× ITS (insulin-transferrin-selenium), hydrocortisone (0.5 μg/ml), and 1× normocin] containing type 3 collagenase (2 mg/ml; Worthington), hyaluronidase (100 U/ml; Sigma) at 37°C until fully digested. Cells were pelleted for 5 min at 300g, resuspended in red blood cell lysis buffer (Abcam) to lyse red blood cells, and pelleted again. Cells were then resuspended in digestion buffer containing deoxyribonuclease 1 (DNase 1) (200μg/ml; Sigma) and incubated for 10 min at 37°C. After DNase digestion, cells were pelleted, resuspended in media, and plated. Cells were grown in modified M87 media containing 2% FBS (50 ). The presence of the NSCLC marker TTF1 (EP1584Y) (1:50, Abcam) and pan-cytokeratin (clone PCK-26) (1:300, Abcam) and the absence of the fibroblast marker S100A4 (EPR2761) (1:100, Abcam) were used to verify the purity of these lines (51 (link)).