description: Protocols: WT and cMKO mice aged 8 and 18 weeks were sacrificed by cervical dislocation and hearts were quickly excised and rinsed with cold sterile PBS. Left ventricle posterior walls were collected and snap frozen in liquid nitrogen and stored at -80°C until use Animals were handled according to the European legislation for animal welfare (Directive 2010/63/EU). All animal protocols were reviewed and approved by the local and national authorities. Mice were housed within a specific pathogen free facility and maintained under standard housing conditions of a 14-10h light-dark cycle, 50-70% humidity, 19-21°C with free access to food and water in cages enriched with bedding material and gnawing sticks. Mtfp1 conditional mice (Mtfp1LoxP/LoxP) were generated by PolyGene AG (Switzerland) on a C57Bl6/N background. Cardiomyocyte specific Mtfp1 KO mice (cMKO; Myh6-Cretg/+Mtfp1LoxP/LoxP) were generated by crossing Mtfp1 conditional mice (Mtfp1LoxP/LoxP) with transgenic (Tg) mice expressing the Cre recombinase under the control of the cardiac alpha myosin heavy chain 6 promoter (Myh6-Cre)(Agah et al., 1997). Littermates that were homozygous for the conditional allele and negative for Myh6-Cre were used as controls (WT; Myh6-Cre+/+Mtfp1LoxP/LoxP).  Heterozygous whole-body Mtfp1 KO mice (Mtfp1+/-) were generated by crossing Mtfp1 conditional mice (Mtfp1LoxP/LoxP) with transgenic (Tg) mice expressing the Cre recombinase under the control of the CMV promoter (CMV-Cre)(Agah et al., 1997) and backcrossing to C57Bl6/N wild type (WT) mice. Cardiomyocyte specific Flag-MTFP1 Knock-In (KI) mice (cMKI; Myh6-Cretg/+Mtfp1+/+,, CAGttg+/) were generated by crossing an inducible mouse model for mCherry-P2A-FLAG-MTFP1 generated by PolyGene AG (Switzerland) on a C57Bl6/N background expression under the CAG promoter with mice expressing the Cre recombinase under the control of the cardiac alpha-(α) myosin heavy chain 6 promoter (Myh6-Cre). Total RNA was extracted by using TRIzol (Invitrogen, NY, USA) according to standard procedures. The Trizol/chloroform mixture was centrifuged at 12000g for 15 min at 4°C, the supernatant corresponding to the aqueous phase was transferred to a new tube, mixed with ethanol 70%, and then applied to an RNA mini-column extraction kit (NucleoSpin RNA kit, MACHEREY-NAGEL) according to the manufacturer's recommendations. After DNA digestion, total RNA was eluted with RNase-free water and stored in liquid nitrogen. RNA was quantified by using the Bioanalyzer 2100 (Agilent) and only RNA samples (≥ 50 ng/µL) with an RNA integrity value (RIN) ≥ 6.8, OD260/280 ≥ 2, OD260/230 ≥ 2 were considered for the transcriptome profiling For RNA sequencing, libraries were built using a TruSeq Stranded mRNA library Preparation Kit (Illumina, USA) following the manufacturer's protocol.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2021-11-25
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2021-11-25
attributes.tag: External Id
attributes.value: SAMEA10289497
attributes.tag: INSDC center alias
attributes.value: 1. Plate-forme Technologique Biomics – Centre de Ressources et Recherches Technologiques (C2RT), Institut Pasteur, Paris, France. 2. Hub de Bioinformatique et Biostatistique - Departement Biologie Computationnelle, Institut Pasteur, 75015 Paris, France
attributes.tag: INSDC center name
attributes.value: 1. Plate-forme Technologique Biomics – Centre de Ressources et Recherches Technologiques (C2RT), Institut Pasteur, Paris, France. 2. Hub de Bioinformatique et Biostatistique - Departement Biologie Computationnelle, Institut Pasteur, 75015 Paris, France
attributes.tag: INSDC first public
attributes.value: 2021-11-25T13:02:36Z
attributes.tag: INSDC last update
attributes.value: 2021-11-25T13:02:36Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-11003:WT5
attributes.tag: age
attributes.value: 18
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: house mouse
attributes.tag: disease
attributes.value: normal
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: individual
attributes.value: WT5
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: heart
attributes.tag: sample name
attributes.value: E-MTAB-11003:WT5