*2.7. Quantitative Real-Time PCR*

RNA extraction from full liver lysates and elimination of genomic DNA was performed using the RNeasy Mini- (QIAGEN, Hilden, Germany) and TURBO DNAfree-Kit (Thermo Fisher Scientific, Waltham, MA, USA), each following the manufacturer's instructions. RNA integrity and purity were assessed by gel electrophoresis and spectrophotometry, equal amounts of RNA were then subjected to cDNA synthesis, using the iScript cDNA Synthesis-Kit (Bio-Rad, Hercules, CA, USA). qPCR was carried out, including one of the primer pairs listed in Supplementary Table S2 and SYBR-Green/ROX dye. *Hprt* was validated and used as a reference gene. Statistical tests and computation of confidence intervals were performed on ΔCT-values, calculated as

$$
\Delta \mathbf{C}\_T = \mathbf{C}\_T \text{(reference gene)} - \mathbf{C}\_T \text{(gene of interest)}.\tag{1}
$$

Fold-changes were calculated as

$$\text{fold-charge} = 2^{\Lambda \text{Cyl}(\text{FC} - \text{Abl.}) - \Lambda \text{Cyl}(\text{Crl})}.\tag{2}$$
