Cx3cr1 13885 1 ap

Total RNA was isolated from mouse liver tissue and cDNA was synthesized using HiScript II Reverse Transcriptase (Vazyme, Nanjing, China). Real-time quantitative polymerase chain reaction (RT-qPCR) analysis was performed using SYBR Green mixture (Vazyme Biotech, Q711). We set actin as the reference gene for each sample. All primers were purchased from Sangon Biotech (Shanghai, China). The primers are given in Supplementary Table 1. For western blot (WB) analysis, total proteins were extracted from the liver samples with lysis buffer (ThermoFisher, 78443) containing 1% PMSF. Proteins (15μg) of each sample were loaded onto a gradient SDS-PAGE. Proteins were then transferred to a PVDF membrane. After blocking with 1% bovine serum albumin (Sigma, A7030), the primary antibodies were used for detection: MAFB (TD8895, 1:2000) (Abmart), CX3CR1 (13885-1-AP, 1:1000) (Proteintech) and Actin (66009-1-Ig, 1. 10000). Tissue levels of MAFB and CX3CR1 were also analyzed by immunohistochemistry (IHC) using MAFB (TD8895, 1:500) (Abmart) and CX3CR1 (13885-1-AP, 1:200) (Proteintech) according to standard protocols.