Liver ChIP Assay for FXR and RXRα Binding

Liver ChIP assay was performed as described previously (Jung et al., 2020 (link); Seok et al., 2018 (link)). Briefly, chromatin extracts were prepared from Fxr-Flox and Fxr-LKO mouse livers after treatment of the mice with GW4064 which was followed by preclearing and immunoprecipitation using control IgG or FXR antibody (Novus Biologicals, NBP2-16550; Santa Cruz, sc-25309), RXRα antibody (Proteintech, catalog no. 21218-1-AP), or BRD4 antibody (Bethyl Laboratories, catalog #A301-985A50). Enrichment in chromatin precipitates of gene sequences was measured by qPCR using primers listed (Supplementary file 5).

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Chen J., Wang R., Xiong F., Sun H., Kemper B., Li W, & Kemper J. (2024). Hammerhead-type FXR agonists induce an enhancer RNA Fincor that ameliorates nonalcoholic steatohepatitis in mice. eLife, 13, RP91438.

Publication 2024

NBP2-16550 RXRα antibody BRD4 antibody

Corresponding Organization :

Other organizations : University of Illinois Urbana-Champaign, The University of Texas Health Science Center at Houston

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Variable analysis

independent variables

Treatment of the mice with GW4064

dependent variables

Enrichment in chromatin precipitates of gene sequences measured by qPCR

control variables

Chromatin extracts prepared from Fxr-Flox and Fxr-LKO mouse livers
Preclearing and immunoprecipitation using control IgG or FXR antibody, RXRα antibody, or BRD4 antibody

controls

Positive control: Immunoprecipitation using FXR antibody
Negative control: Immunoprecipitation using control IgG

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