Fibroblasts were isolated from P5 and P12 mouse lungs using previously described methods (12). Briefly, after perfusion with sterile PBS, mouse lungs were harvested, dissected, and enzymatically digested in DMEM containing collagenase type I (0.5 mg/mL; Sigma-Aldrich) and collagenase type IA (0.5 mg/mL; Sigma-Aldrich), followed by trituration to mechanically dissociate lung tissue. The resulting cell suspension was sequentially filtered through 100, 70, and 20 μm sterile filters. The filtrate from the 20 μm filter was plated in 100 mm cell-culture dishes and cultured in DMEM supplemented with 10% FCS. For all experiments, passage 3 or 4 fibroblasts were first plated in 12- or 24-well plates or chamber slides and cultured for 72 hours. After overnight serum starvation, cells were exposed to various treatment conditions for 4–48 hours in serum-free conditions prior to harvesting for RNA extraction or protein quantification.

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