description: Protocols: Murine tissues for protein or RNA extraction were frozen at the time of collection and stored at -80. All data are from male mice. Insig1 KO mice were generated on a C57Bl6/J background and compared to wild-type and heterozygous littermates. Mice were maintained in a room with controlled temperature (21°C; 12 hr light/dark cycle) with free access to food and water; the pathogen-free facilities were compliant with the FELASA Guidelines. The UK Home Office and the USC Bioethics Committee of the Universities of Cambridge and Newcastle approved all animal procedures. Mice were fed on a chow diet (Safe Diets, Code ds-105) until they were enrolled in specific procedures. CCL4 and VOO (Virgin Olive Oil) Animals; 12 week old Insig1 KO, Het, or WT underwent intraperitoneal (IP) injection of 2 ul/BW(g) of the CCl4: virgin olive oil (1:1 (v/v)) mix or virgin olive oil alone. Mice were humanely culled under terminal isofluorane anaesthesia four days after the IP injection, by cutting the heart and inferior vena cava. Blood was collected from the chest cavity. These experiments were approved by the Animal Welfare and Ethical Review Board and carried out at Newcastle University Comparative Biology Centre under a UK Home office license. Murine RNA was isolated using miRNAeasy Mini Kits (Qiagen), according to the manufacturer's instructions. All reagents, plastic ware, and supplies used were nuclease-free, sterile, and of molecular biology grade. RNA purity (A260/A280>1.80) and concentration were determined using the Nanodrop spectrophotometer (Thermo Fisher Scientific, Delaware USA). RNA integrity was studied using the 2100 Bioanalyzer (Agilent) and RNA 6000 Nano Kits (Agilent, Santa Clara, California, USA). RNA Integrity Number (RIN) of 8 was considered the lowest cut-off for RNA sequencing. RNA from tissues (1 ug RNA) were used to generate barcoded sequencing libraries using Illumina TruSeq® Stranded mRNA Library Preparation Kit (Illumina) following manufacturer's instructions. Library preparation was performed by the Genomics and Transcriptomic Core at the Institute of Metabolic Science.
attributes.tag: ENA first public
attributes.value: 2021-04-15
attributes.tag: ENA last update
attributes.value: 2020-12-09
attributes.tag: External Id
attributes.value: SAMEA7699107
attributes.tag: INSDC center alias
attributes.value: Co-author
attributes.tag: INSDC center name
attributes.value: Co-author
attributes.tag: INSDC first public
attributes.value: 2021-04-15T00:32:17Z
attributes.tag: INSDC last update
attributes.value: 2020-12-09T12:53:31Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-9864:Sample 24
attributes.tag: age
attributes.value: 12
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: diet
attributes.value: virgin olive oil
attributes.tag: disease
attributes.value: normal
attributes.tag: duration of experiment
attributes.value: 0.5
attributes.tag: genotype
attributes.value: Insig1 +/-
attributes.tag: individual
attributes.value: D712_D508_VOO
attributes.tag: organism part
attributes.value: liver
attributes.tag: sample name
attributes.value: E-MTAB-9864:Sample 24
attributes.tag: sex
attributes.value: male
attributes.tag: strain
attributes.value: C57Bl6/J