Tissues were mechanically disrupted and simultaneously homogenized in the presence of QIAzol Lysis reagent (Qiagen, Valencia, CA, USA), using a Mikrodismembrator (Braun Biotech International, Melsungen, Germany). RNA was extracted using the miRNeasy Mini kit (Qiagen) according to manufacturer’s instructions. RNA concentrations were measured with the NanoDrop ND-100 Spectrophotometer (NanoDrop Technologies, Wilmington, DE) while RNA quality was assessed with the Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA) using the RNA 6000 Nano kit (Agilent Technologies). Samples, included in the present analysis, had a RIN score greater than 5 and a 28S:18S rRNA ratio near to 2:1.
Label
Hy3
Label protocol
Total RNA from each sample was labeled using miRCURY labelling Exiqon kit (Exiqon, Vedbaek, Denmark)
Hybridization protocol
Labeled RNA was hybridized using the GeneTaq Hybridization Station and slides were manually washed
Scan protocol
Fluorescent signals were detected using a GenePix4100 scanner
