Total RNA was prepared from snap-frozen male and female adrenal and pancreatic tissue using Qiagen RNeasy Lipid Tissue Mini Kit Cat # 74804 (Qiagen, CA, USA) according to the manufacturer’s instructions, and stored at − 80 o C, as described previously [35 (link)]. This method was slightly modified for pancreatic RNA extraction, according to De Lisle, 2014 [36 ]. RNA integrity was measured using a 2100 Bioanalyzer instrument and an RNA 6000 Nano LabChip assay (Agilent Technologies, CA, USA). RNA concentrations were determined by absorption at 260-nm wavelength with an ND-8000 spectrophotometer (Nanodrop Technologies, DE, USA).
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