For chromatin immunoprecipitation (ChIP), 5 month-old male C57BL/6J mice were euthanized by CO2 asphyxiation at ZT 8. Livers were removed and pooled for ChIP, processing and sequencing. Tamoxifen-induction of Cre Recombinase activity was accomplished by daily intraperitoneal injection of 2 mg tamoxifen (Sigma) in 100μl corn oil (Sigma) for 7 days. 3 month-old tamoxifen-treated animals were subjected to wheel running assays for 7 days after the end of treatment. For gene expression analysis, 5–6 month-old males were euthanized by cervical dislocation at indicated ZT points for rapid dissection and snap-freezing of the tissues. During the dark cycle, procedures were performed under red light. Detailed materials and methods are provided in the Supplementary Information .
Protocol detail
Circadian Regulation via ChIP-seq
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Animals
Asphyxiation
Assays
C57bl mice
Cervical
Chromatin immunoprecipitation
Corn oil
Cre recombinase
Dislocation
Dissection
Gene expression analysis
Intraperitoneal injection
Light
Livers
Males
Tamoxifen
Tissues
Corresponding Organization : Howard Hughes Medical Institute
Other organizations : University of California, San Diego, University of Sydney, Westmead Institute, Westmead Hospital, École Polytechnique Fédérale de Lausanne
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Variable analysis
independent variables
- Tamoxifen-induction of Cre Recombinase activity accomplished by daily intraperitoneal injection of 2 mg tamoxifen (Sigma) in 100μl corn oil (Sigma) for 7 days
- Wheel running assays for 7 days after the end of treatment
- Gene expression analysis
- 5 month-old male C57BL/6J mice
- Liver tissue
- Euthanasia by CO2 asphyxiation at ZT 8
- Rapid dissection and snap-freezing of tissues
- Procedures performed under red light during the dark cycle
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