Itgb1

Flow cytometry was performed as described [12 (link)]. Following antibodies were used: ITGAV (327,907, BioLegend), ITGB1 (11–0299, Thermo Fisher Scientific), ITGB3 (336,403, BioLegend), ITGB5 (11–0497-42, Thermo Fisher Scientific), ITGB6 (FAB4155P, R&D), HLA-DR (347,401, BD). Stained cells were subjected to FACS Calibur Flow Cytometry System (BD).

TaqMan qRT-PCR probes and primers were obtained from Thermo Fisher Scientific (Waltham, MA, USA) as follows: miR-124-3p (product ID: 000446; Thermo), ITGA3 (product ID: Hs01076879_m1), ITGB1 (product ID: Hs01127536_m1) and ITGA2 (product ID: Hs00158127_m1). GUSB (product ID: Hs99999908_m1) and RNU48 (product ID: 001006) were used as internal controls. The procedure for qRT-PCR was described in previous studies [30 (link), 41 (link)].

Protein lysates, collected in 9 mol/L urea and 0.075 mol/L Tris, pH 7.6, were quantified using a Bradford protein quantification assay. Samples were normalized for protein concentration. Protein visualization was performed using a Jess Automated Protein Analysis system (BioTechne, Minneapolis, MN, USA). Antibody dilutions were as follows: DDR2 (1:50, Cell Signaling Technology, Danvers, MA, USA), POSTN (1:200 Adipo-gen, San Diego, CA, USA), pAKT (1:50, Cell Signaling Technology, Danvers, MA, USA), pSrc (1:50, Cell Signaling Technology, Danvers, MA, USA), ITGB1 (1:50, Thermofisher, Waltham, MA, USA), Actin (1:200, Sigma-Aldrich, Burlington, MA, USA). For signaling pathway analysis, CAFs were embedded in 1 mg/mL polymerized collagen 1 overnight prior to collecting protein lysates.