CyTOF of mouse liver transplantation samples from FGF21 KO and wild-type mice

To elucidate the underlying mechanism of inflammatory injury driven by FGF21 deficiency, we performed CyTOF of mouse liver transplantation samples from FGF21 KO and wild-type mice. Transplanted livers from 8 mouses (n = 4/group) were analysed by CyTOF. CyTOF analyses were performed by PLTTech Inc. (Hangzhou, China). Briefly, mouse liver tissue was dissociated into single cells using DNase, collagenase IV and hyaluronidase (Sigma-Aldrich, St. Louis, MO, USA). Immune cells were enriched using Percoll density gradient media (Sigma-Aldrich), and red blood cells were removed using ACK Lysing Buffer (Sigma-Aldrich). Qualified samples were blocked and stained for 30 min with a panel of 42 antibodies (Supplementary Table 1), followed by fixation overnight. Permeabilization buffer was applied, and the cells were incubated in an intracellular antibody mix. The cells were rinsed, and the signals were detected using a CyTOF system (Helios, Fluidigm, CA, USA). The types of immune cells were identified via nonlinear dimensionality reduction [t-distributed stochastic neighbor embedding (tSNE)], followed by density clustering.