Total RNA was extracted from neonatal lungs and lung fibroblasts using TRIzol reagent (Life Science Technologies) using standard methods (67). First-strand cDNA was synthesized using oligo-dT primers and Moloney murine leukemia virus reverse transcriptase (Superscript II; Invitrogen). PCR primers were designed using PrimerQuest design tool (Integrated DNA Technologies). Two-step PCR was performed using an Applied Biosystems StepOne-Plus Real Time PCR system (Applied Biosystems) and SYBR green detection system (Bio-Rad). Gene expression was normalized to β-actin in each sample. Fold change in mRNA expression was calculated using the 2-ddCT method (68).

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