NextSeq 500 paired end sequencing; GSM3435447: Sample-1; Mus musculus; RNA-Seq

Sample-1

We provide here the alterations in gene expression profiles of 3T3-L1 cells, a validated model for adipogenesis, exposed to quizalofop-p-ethyl for 6h, 24h and 12 days. Overall design: Exposure to endocrine disrupting chemicals is a risk factor for obesity. The most commonly used pesticide active ingredients have never been tested in an adipogenesis assay. We tested for the first time the lipid accumulation induced by glyphosate, 2,4-dichlorophenoxyacetic acid, dicamba, mesotrione, isoxaflutole and quizalofop-p-ethyl (QpE) in 3T3-L1 adipocytes. Only QpE caused triglyceride accumulation from a concentration of 1 µM. We thus conducted an in-depth investigation of molecular mechanisms responsible for the adipogenic effects of quizalopfop-p-ethyl by an RNA-seq analysis.

Quizalofop-p-ethyl induces adipogenesis in 3T3-L1 cells

Submitted on 19-APR-2019

We provide here the alterations in gene expression profiles of 3T3-L1 cells, a validated model for adipogenesis, exposed to quizalofop-p-ethyl for 6h, 24h and 12 days. Overall design: Exposure to endocrine disrupting chemicals is a risk factor for obesity. The most commonly used pesticide active ingredients have never been tested in an adipogenesis assay. We tested for the first time the lipid accumulation induced by glyphosate, 2,4-dichlorophenoxyacetic acid, dicamba, mesotrione, isoxaflutole and quizalofop-p-ethyl (QpE) in 3T3-L1 adipocytes. Only QpE caused triglyceride accumulation from a concentration of 1 ÂµM. We thus conducted an in-depth investigation of molecular mechanisms responsible for the adipogenic effects of quizalopfop-p-ethyl by an RNA-seq analysis.

RNA extraction was performed using the Qiagen RNeasy kit (Qiagen, Manchester, UK) according to the manufacturer's instructions. RNA samples were assessed for quantity and integrity using the NanoDrop 8000 spectrophotometer V2.0 (ThermoScientific, USA) and Agilent 2100 Bioanalyser (Agilent Technologies, Waldbronn, Germany), respectively. All samples displayed a RIN score of 10 and were taken forward for RNA library preparation. A 100ng aliquot of total RNA from each sample was used to prepare mRNA libraries using the NEBNext mRNA isolation kit in conjunction with the NEBNext Ultra II Directional RNA Library preparation kit (New England Biolabs, Massachusetts, USA), and samples randomised before preparation. Fragmentation of isolated mRNA prior to first strand cDNA synthesis was conducted using incubation conditions recommended by the manufacturer for a fragment size of 300bp (94°C for 10 minutes). A total of 13 cycles of PCR were performed for final library amplification. Resulting libraries were quantified using the Qubit 2.0 spectrophotometer (Life Technologies, California, USA) and average fragment size assessed using the Agilent 2200 Tapestation (Agilent Technologies, Waldbronn, Germany). A final sequencing pool was created using equimolar quantities of each sample with compatible indexes. Paired-end reads of 75bp were generated for each library using the Illumina NextSeq®500 in conjunction with the NextSeq®500 v2 High-output 150-cycle kit (Illumina Inc., Cambridge, UK).

Quizalofop-p-ethyl induces adipogenesis in 3T3-L1 cells

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