Sc 20806

Procedure is detailed in supplementary information. The slides were incubated at 4°C overnight with primary antibody included anti-NTS (1:200, SC-20806, Santa Cruz biotechnology®), and anti-ErbB3 (1:50, NCL-c-erbB-3, Novocastra™), anti-NTSR1 (1:100; SC-7596, Santa Cruz Biotechnology®) and anti-ErbB2 (1:400, A0485, Dako) was incubated at room temperature for 1hour and 30 minutes respectively. The levels of staining were scored based on staining intensity within the tumor sample, with weak as 1, moderate as 2, strong as 3.

Procedure is detailed in SI. For all cases histologic slides of primary tumors were obtained from paraffin wax embedded tissues. Standard H&E staining was used to ensure the tumoral character of the specimen. Deparaffinized tissue sections (4 μm) were incubated at 4°C overnight with primary antibody included anti-NTS (1:200, SC-20806, Santa Cruz biotechnology®), anti-NTSR1 (1:100; C-20, Santa Cruz Biotechnology®) and anti-ErbB3 (1:50, NCL-c-erbB-3, Novocastra™), and anti-ErbB2 (1:400, A0485, Dako) was incubated at room temperature for 30 minutes.
For prognosis evaluation, all specimens were scored by an anatomopathologist with special interest in pulmonary pathology (DD). NTSR1 staining of cancer cells was scored as positive in the presence of staining cells > 10 %. Semi-quantitative evaluation was also performed: 0: no staining; 2: more than 50% of tumor cell showing a positive stain of high intensity; 1: intermediate cases.