Rev erbα

Mice livers or primary murine hepatocytes were subject to crosslinking in 1% of formaldehyde for 5 min followed by quenching with glycine for 5 min on ice. Cells were pelleted by centrifugation and resuspended in a lysis buffer (50 mM HEPES pH 8.0, 140 mM NaCl, 1 mM EDTA, 10% glycerol, 0.5% NP40, 0.25% Triton X100). The pellets were then resuspended in a washing buffer (10 mM Tris pH 8.0, 1 mM EDTA, 0.5 mM EGTA, 200 mM NaCl), washed and resuspended in a shearing buffer (0.1% SDS, 1 mM EDTA, pH 8, 10 mM Tris HCl, pH 8) before sonication following the instruction of the manufacturer. Chromatin fragments were precipitated using specific antibodies and protein G beads, washed, and treated with proteinase K and RNase A. Purified chromatin immunoprecipitation (ChIP) DNA was then used for chromatin immunoprecipitation quantitative real-time PCR (ChIP-qPCR) analysis. The antibodies used for the ChIP-qPCR assay include PPARα (Millipore, MA, USA; MAB3890); RNAPII (Santa Cruz Biotechnology, TX, USA; sc-899); RNAPII-S2P (Active Motif, CA, USA; #61083); H3K27ac (Abcam, Cambridge, UK; ab4729); H3K4me1(Abcam, Cambridge, UK; ab8895); IgG (Santa Cruz Biotechnology, TX, USA; sc-2027), and REV-ERBα (Proteintech, IL, USA; 14506-1-AP). ChIPs were performed with each experimental point in triplicate, and each experiment was repeated three times. The primers are shown in Supplementary Table 1.

Primary antibodies Rev-erbα, Bmal1 and Beclin1 were purchased from Proteintech Wu Han China. Primary antibodies LC3 and P62 were purchased from Cell Signaling Technology USA. Primary antibodies Pparα and Srebp1c were purchased from Affinity Biosciences USA. β-Actin, secondary antibodies and goat anti-mouse Alexa Fluor 488 were purchased from Zhongqiao Jinshan China. Nucleoprotein/cytoplasmic protein extraction kit was purchased from Best Bio Shanghai, China. GSK4112 (CAS number: 1216744–19-2) was purchased from TargetMol China. SR8278 (CAS number: 1254944-66-5) was purchased from APExBIO USA. ORO was purchased from Sigma USA. Rev-erbα expression interference plasmid and Control plasmid was purchased from GenePharma (ShangHai, China). Bmal1 small interfering RNA (siRNA) and Control siRNA were synthesized by GenePharma (Shanghai, China).