Protein l agarose

Formaldehyde cross‐linked chromatin complexes were immunoprecipitated using an antibody against C/EBPα or without an antibody (as the negative control) and collected by incubation with Protein L—agarose (Santa Cruz Biotechnology, Dallas, Texas, USA). The DNA‐protein cross‐links were then reversed by heating. The DNA was purified and subjected to PCR and RT‐qPCR using primers covering the C/EBPα binding site on the miR‐128 promoter. Primers used for PCR are listed in Table S2.

Small intestine tissue was collected from WT mice (no flox). Intestinal protein supernatants were prepared and the protein concentration was assayed as described above. The supernatants were immunoprecipitated with a polyclonal CIDEC antibody (Cloud-Clone Corp; Wuhan, China) following the manufacturer's protocol. Briefly, 3 mg protein supernatant samples were precleared by IgG and arotein L-agarose (Santa Cruz Biotechnology; Dallas, TX, USA) for 30 min, incubated with 2 mg of primary antibody for 1h and then incubated with protein L-agarose at 4 °C overnight (16 h). The bound proteins were washed twice with prechilled RIPA and eluted by boiling in Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) loading buffer (Solarbio; Beijing, China). The resulting proteins were assayed by SDS-PAGE and sent to APTBIO (Shanghai, China) for mass spectrum analysis.