Total RNA from homogenized lung was extracted using TRIzol (Invitrogen, Carlsbad, CA), and reverse transcribed using the SuperScript III First-Strand Synthesis System for RT-PCR (Invitrogen). RT-PCR for PDGFR-α (primer from Thermo Fisher Scientific Cat number: 4331182 Assay ID: Hs00998018_m1) and miR-219-5p (primer from Thermo Fisher Scientific hsa-miR-219 Cat number: 4427975 Assay ID 000522) on homogenized lung were performed as described previously [23] using specific primers. Briefly, for RT of mature miRNA, 100 ng of DNase-treated total RNA was reverse transcribed using 100 U of SuperScript III Reverse Transcriptase enzyme and 100 nM of miR-219-5p stem-loop oligonucleotides [24]. The cDNA samples were then used for real-time PCR using miRNA-specific primers for miR-219-5p. Real-time PCR was performed on the MyiQ™ Single-Color Real-Time PCR detection System (Bio-Rad) using SYBR Green PCR Master Mix (Applied Biosystems). Two microliters of each cDNA sample were subjected to real-time PCR in a total volume of 20 µl using 100 nM of each primer. Thermocycling of miRNA cDNA was performing after 10 min of initial denaturation at 95 °C followed by 50 cycles of 15-s at 95 °C and 1 min annealing/extension at 60 °C. miR-219-5p expression levels were normalized to U6 snRNA. Expression levels of genes were normalized to 18S rRNA.

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