*2.4. Sample Collection*

At the time points indicated in the result section, the mice were anaesthetised by an i.p. injection of ketamine (100 mg/kg b.w.) and xylazine (10 mg/kg b.w.). After the loss of reflexes, blood as well as liver tissue samples were collected. The blood samples were collected from the portal vein, the hepatic vein, and the right heart chamber in EDTA-coated syringes as previously described [15]. The samples were centrifuged at 13,000 rpm for 10 min in order to separate plasma. The collected plasma was stored at −80 ◦C until used for analyses. After blood collection, the remaining blood was removed by perfusion trans-cardially with 40 mL PBS. Subsequently, the whole liver was excised and specimens were collected from defined anatomical positions as follow: (i) a specimen of approximately 1 cm size was taken from the left liver lobe, fixed in 4% paraformaldehyde (PFA) for 2 days and then embedded in paraffin for 2D staining; (ii) a specimen of approximately 0.5 cm size was taken from the left liver lobe and immediately embedded in Tissue-Tek®cryomold in Neg-50 media (ThermoFisher Scientific, Oberhausen, Germany), frozen in 2-methylbutane and stored at −80 ◦C until used for cryosection preparation; (iii) a specimen of approximately 0.5 cm size was taken from the median liver lobe, fixed in 4% PFA for 2 days, incubated in 30% sucrose for 2 days, and then embedded in Tissue-Tek®cryomold in Neg-50 media, frozen in 2-methylbutane and stored at −80 ◦C until used for preparation of liver slices; (iv) a specimen of approximately 20 mg weight was taken from the left liver lobe, snap-frozen in liquid nitrogen and stored at −80 ◦C until used for RNA isolation.
