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Mouse Embryonic Fibroblasts (MEFs) Adipogenic Potential: A Comprehensive Transcriptome Analysis

ABSTRACT: Mouse Embryonic Fibroblasts (MEFs) Adipogenic Potential: A Comprehensive Transcriptome Analysis

PROVIDER: PRJNA640338 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR12044367_1.fastq.gz	Fastqsanger.gz
	SRR12044367_2.fastq.gz	Fastqsanger.gz
	SRR12044368_1.fastq.gz	Fastqsanger.gz
	SRR12044368_2.fastq.gz	Fastqsanger.gz
	SRR12044369_1.fastq.gz	Fastqsanger.gz

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Mouse Embryonic Fibroblasts (MEFs) Adipogenic Potential: A Comprehensive Transcriptome Analysis

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RNAseq from Mfn1/2 knockout MEFs with adipogenic differentiation

Project description:Objective: A biallelic missense mutation in mitofusin 2 (MFN2) causes multiple symmetric lipomatosis and partial lipodystrophy, implicating disruption of mitochondrial fusion or interaction with other organelles in adipocyte differentiation, growth and/or survival. In this study, we aimed to document the impact of loss of mitofusin 1 (Mfn1) or 2 (Mfn2) on adipogenesis in cultured cells. Methods: We characterised adipocyte differentiation of wildtype (WT), Mfn1-/- and Mfn2-/- mouse embryonic fibroblasts (MEFs) and 3T3-L1 preadipocytes in which Mfn1 or 2 levels were reduced using siRNA. Results: Mfn1-/- MEFs displayed striking fragmentation of the mitochondrial network, with surprisingly enhanced propensity to differentiate into adipocytes, as assessed by lipid accumulation, expression of adipocyte markers (Plin1, Fabp4, Glut4, Adipoq), and insulin-stimulated glucose uptake. RNA sequencing revealed a corresponding pro-adipogenic transcriptional profile including Pparg upregulation. Mfn2-/- MEFs also had a disrupted mitochondrial morphology, but in contrast to Mfn1-/- MEFs they showed reduced expression of adipocyte markers. Mfn1 and Mfn2 siRNA mediated knockdown studies in 3T3-L1 adipocytes generally replicated these findings. Conclusions: Loss of Mfn1 but not Mfn2 in cultured pre-adipocyte models is pro-adipogenic. This suggests distinct, non-redundant roles for the two mitofusin orthologues in adipocyte differentiation.

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Mus musculus strain:mouse embryonic fibroblasts (MEFs) | isolate:MEFs | breed:MEFs | cultivar:Mus musculus

Project description:Mus musculus strain:mouse embryonic fibroblasts (MEFs) | isolate:MEFs | breed:MEFs | cultivar:Mus musculus Raw sequence reads

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Mus musculus strain:mouse embryonic fibroblasts (MEFs) | isolate:MEFs | breed:MEFs | cultivar:Mus musculus

Project description:Mus musculus strain:mouse embryonic fibroblasts (MEFs) | isolate:MEFs | breed:MEFs | cultivar:Mus musculus Raw sequence reads

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Expression analysis of mouse embryonic fibroblasts (MEFs) and Thymidine treatment-induced MEFs (induced cells)

Project description:Investigation of differentially expressed genes in thymidine treatment-induced MEFs. Mouse embryonic fibroblasts (MEFs) were obtained at E14.5 embryos and supplied with Dulbecco’s modified Eagle’s medium (DMEM) (Invitrogen) with 15% FBS, 100 μg/ml streptomycin and 100 U/ml penicillin. Potential hematopoietic progenitor cells (CD45+CD41+CD31+c-Kit+) were negatively selected with Biotin labeled anti-CD45, anti-CD31, anti-CD41 and anti-c-Kit antibodies by magnetic beads. Hereafter, treated MEFs are referred to as 4neg MEFs for initiation of induction. 4neg MEFs were seeded on mitomycin (MMC)-treated MEFs as feeder cells and cultured in DMEM with 15% FBS, 100 μg/ml streptomycin, 100 U/ml penicillin, 2 mM L-glutamine, 0.1 mM non-essential amino acids and 100 μM thymidine as induction medium for 6 days. Cells were then trypsinized and re-seeded to new feeder cells for another 6 days. Cells were positively selected with Biotin labeled anti-CD45 by magnetic beads followed with analysis.

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