Anti klf15 antibody

The ChIP assay was carried out using the anti-KLF15 antibody, as described previously^{11 (link),18 (link)}. Briefly, soluble chromatin was subjected to immunoprecipitation using the anti-KLF15 antibody (SantaCruz Biotechnology). After recovering DNA, the purified DNA samples were utilized for qPCR with specific primers covering the (proximal and distal regions) of the Fgf21 promoter. The primers used for PCR were as follows: proximal region, forward 5′-GGGTTCCTCCTAGAAATCCA-3′ and reverse 5′-CAGTCACCCTCACCAACCCC-3′; distal region, forward 5′-GCTGCGCCCTCTCCTCCGCC-3′ and reverse 5′-TGGGAACGT-GCATAGAACCT-3′.

ChIP assay was done as described 27 (link), 29 (link) using the Thermo ChIP Kit (Invitrogen). Cells (2x10⁶) were crosslinked with a 1% formaldehyde solution for 10 min and terminated by adding lycine 1.25 M). After being washed twice using ice-cold PBS, cells were harvested in cell scraper and then lysed in 200 μL of SDS lysis buffer, and sonicated to generate 300 to 800 bp DNA fragments. The lysate was pre-cleared by incubation with protein G agarose and incubated overnight at 4 °C with either anti-KLF15 antibody (Santa Cruz Biotechnology) or non-immune IgG (Upstate Biotechnology, Inc.). To collect the immunoprecipitated complexes, protein G magnetic beads were incubated. After purified, DNA samples were amplified in a 7500 quantitative real-time PCR System (Applied Biosystems, Carlsbad, CA) and quantitated in triplicate by SYBR Green qPCR (Takara Biotechnology, Dalian, China) using forward and reverse primer sequences (Supplementary Table S3) for the mouse flot2 promoter. Data were analyzed using the 2^-△△CT method.