RT-PCR
Equivalent amounts of purified RNA obtained from neutrophils, leukocytes, and brain tissue were used as a template for cDNA synthesis using SYBR Green qPCR Master Mix (Fermentas, Burlington, Canada), oligo-d(T) primers, and SuperScript III/RNaseOUT Enzyme Mix (Invitrogen, Carlsbad, CA, USA). MiR-29b primers for human neutrophils were 5’-GGGTAGCACCATTTGAAATCA3’ and 5’-GTGCGTGTCGTGGAGTCG-3’. MiR-29b primers for rat leukocytes were 5’-CTCAACTGGTGTCGTGGAGTCGGCAATTCAGTTGAG-3’ and 5’-ACACTCCAGCTGGGTAGCACCATTTGAAATC-3’. H19 primers for human neutrophils were 5’-CTTCTTTAAGTCCGTCTCGTTC-3’ and 5’-GAGGCAGGTAGTGTAGTGGTTC-3’. H19 primers for rat 5’- GGAATCGGCTCGAAGGTAAA-3’ and 5’-GGGCCAGGCAGAGTTAGTTG-3’. C1QTNF6 primers for human neutrophils were 5’-TCAGTCCCTTCCACCAAA-3’ and 5’-ACCTTGATAAAGCCTGGAGA-3’. C1QTNF6 primers for rat were 5’-GTTCGGGGTCTGTGAGTTGAG-3’ and 5’-CTTTCAGGATGGTGATGTTGATGTA-3’. The relative gene expression was calculated using the 2-△△CT method, normalised, and expressed as fold change relative to U6.