*2.6. Gene Expression Analyses*

Quantitative real-time polymerase chain reaction (qRT-PCR). Total RNA was isolated from frozen liver tissue using QIAzol Lysis Reagent (Qiagen, Hilden, Germany). cDNA synthesis was performed from 2 μg of isolated RNA using a commercially available kit (High-Capacity cDNA Reverse Transcription Kit, Applied Biosystems, Schwerte, Germany). qRT-PCR analyses were performed using TaqMan 7500 Real-Time PCR, TaqMan universal PCR Master Mix (Applied Biosystems, Schwerte, Germany), and TaqMan gene expression assays (ThermoFisher Scientifics, Oberhausen, Germany). Gene expression values were normalized to GAPDH, calculated using the ΔΔCt method, and are expressed as fold changes over untreated control samples.
