As described previously (16 (link)), murine CAFs were isolated from lung tissues at necropsy by immediately perfusing tissues with 2% fetal bovine serum in Hank’s buffered salt solution (FBS-HBSS) and dispersing them into single cell suspension by immersion in 3 mg/mL of collagenase and Dispase®II (Roche) on a gentleMACS™ Dissociator (Miltenyi Biotec, Bergisch Gladbach, Germany) using the lung tissue dissociation programs (Lung_01 and Lung_02). Dispersed cells were centrifuged, washed with FBS-HBSS, and subjected to red blood cell lysis by adding RBC Buffer (BioLegend). The remaining cells were centrifuged, washed, filtered (70 µm and 40 µm), counted (Countess™, Invitrogen), and mixed twice with antibody-conjugated magnetic beads (Dynal-Magnetic beads, Invitrogen) on a rotator, each time for 45 min at 4°C, to first deplete leukocytes (anti-CD45 and anti-CD68, Abcam), endothelial cells (anti-CD31, BD Pharmingen), and epithelial cells (anti-EPCAM, Pharmingen) and then to isolate fibroblasts (anti-Thy-1, BD Pharmingen) from the supernatants. Fibroblasts were eluted off the anti-Thy-1-conjugated beads by incubation in FBS-HBSS, 0.5% BSA, and 2mM EDTA, centrifuged, washed, and cultured in RPMI1640 containing 10% FBS and 100 mg/100U penicillin-streptomycin (GIBCO).