Immunohistochemistry (IHC) for SCD in Lung Cancer

IHC for SCD was performed in lung adenocarcinoma tissue microarray (US Biomax, Rockville, MD, USA) according to the manufacturer’s instructions [37 (link)]. The prepared tissues microarray slides were incubated with SCD antibody (Cell-Signaling technology) for overnight and incubated with horse anti-rabbit secondary antibody (Vector Laboratories, Burlingame, CA, USA) for 2 h. Following treatment with 3,3’-diaminobenzidine solution (Vector Laboratories), slides were counterstained using hematoxylin, dehydrated in alcohol, and mounted in a xylene-based solution. The mean IHC staining intensity for SCD was quantified using ImageJ software and scored 0–3 as follows [38 (link)]: 0, no staining; 1, < 15 mean intensity; 2, 15–50 mean intensity; and 3, > 50 mean intensity.