do4304
Mus
Protocols: All mice were housed in the same husbandry conditions within the Biological Resources Unit in the Cancer Research UK-Cambridge Institute under a Home Office Licence. C57BL/6J and CAST/EiJ mouse strains were used in experiments as parental strains (F0) as well as for breeding of reciprocal crosses of F1 mice. All mice used in the experiments were males between eight and 12 weeks of age, and harvested at the same time of day (between 8 and 11am). Liver perfusion was performed on mice post mortem, prior to tissue dissection. Harvested tissues were formaldehyde cross-linked for ChIP-seq experiments. Before cross-linking, dissected tissue was immediately chopped post mortem and added to a cross-linking solution containing 1% formaldehyde. Tissue was incubated for 20 min prior to quenching with 1/20th volume of 2.5 M glycine. Samples were incubated for a further 10 min before washing with PBS and flash-freezing and storage at -80oC. To create HNF4A and CEBPA heterozygous knockout mice, we acquired mice with targeted alleles from JAX (www.jax.com) (HNF4A stock number: 004665 (Hayhurst et al. 2001); CEBPA stock number: 006230  (Zhang et al. 2004)). Heterozygous knockouts were generated via the Cre-loxP system (Orban et al. 1992) using the germline deleter strain PgkCre (Lallemand et al. 1998) and crossing it to CebpaFLOX/FLOX and Hnf4aFLOX/WT mice. Ear biopsies were taken at the time of weaning for genotyping to confirm deletion via PCR
isolate
not applicable
organism
Mus
development stage
8 - 12 weeks
genetic modification
floxed allele
genotype
CepbaFLOX/-
individual
34535
organism part
liver
sex
male
strain
mus musculus