Isolation and Culture of CAFs from NSCLC

Primary CAFs used in the present study were isolated from NSCLC specimens, as previously reported [1 (link)]. The study was conducted in accordance with a protocol approved by the University Health Network (UHN) Research Ethics Board. NSCLC tissues were collected with informed consent from all patients. An overview of CAFs used in this study is presented in Table 1. CAF cells were cultured in Dulbecco’s Modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS; Thermo Scientific, Burlington, ON, Canada). All CAF primary cultured cells were maintained and used at early passage (passage 2–5). Murine C2C12 myoblasts stably expressing either human integrin α1 (C2C12-α1), human integrin α2 (C2C12-α2) or human integrin α11 (C2C12-α11) were obtained from Prof. Donald Gullberg (University of Bergen, Norway) and were cultured in DMEM supplemented with 10% FBS. The NCI-H1573, H2073, H358, H1975 and H2009 NSCLC cell lines were obtained from the American Type Culture Collection (Manassas, VA, USA) and were cultured in RPMI 1640 media supplemented with 10% FBS.