RNA Isolation and qPCR Analysis of NF-κB Expression

RNA was isolated from the striatal tissue and qPCR was performed as stated by the instructions from the manufacturer using SV Total RNA Isolation System (Promega Corporation, Madison, WI, USA) and Applied Biosystems StepOne™ (Applied Biosystems, Foster City, CA, USA), correspondingly according to Fathy and Said [40 (link)]. The NanoDrop™ 2000/2000c Spectrophotometer (Thermo Scientific, Lo, UK) was selected for detecting the concentration and the quality of the isolated RNA. NF-кB gene was used with a forward primer sequence: 5′-CCCTACGGAACTGGGCAAAT-3′ and a reverse primer sequence: 5′-GCGGAATCGAAATCCCCTCT-3′ [41 (link)]. NF-кB relative expression level was normalized based on the housekeeping gene “glyceraldehyde 3-phosphate dehydrogenase (GAPDH)” (the sense sequence: 5′-ATGTGTCCGTCGTGGATCTGAC-3′ and the antisense sequence: 5′-AGACAACCTGGTCCTCAGTGTAG-3′) [42 (link)].