For RNA extraction, the tissues were ground and the total RNA was isolated using NucleoSpin® RNA columns (Macherey-Nagel; Düren, Germany) according to the manufacturer’s instructions. RNA was quantified using NanoDrop® spectrophotometer ND-1000 (Thermo Scientific Fisher; Waltham, MA, USA). Total RNA was reverse-transcribed into cDNA with a High Capacity Reverse-Transcription kit (iScriptTM cDNA synthesis KIT; Bio-Rad, Hercules, CA, USA). Real-time PCR detection was performed with a SYBR Green supermix (Bio-Rad, Hercules, CA, USA). Gene specific primers were designed and checked using the BLAST algorithm. Gene expression was normalized using the expression of GAPDH, β-Actin and RPS13.
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