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Next Generation Sequencing Facilitates Quantitative Analysis of MV4-11 cell line Transcriptomes

ABSTRACT: Next Generation Sequencing Facilitates Quantitative Analysis of MV4-11 cell line Transcriptomes

PROVIDER: PRJNA627731 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR11601225_1.fastq.gz	Fastqsanger.gz
	SRR11601225_2.fastq.gz	Fastqsanger.gz
	SRR11601226_1.fastq.gz	Fastqsanger.gz
	SRR11601226_2.fastq.gz	Fastqsanger.gz
	SRR11601227_1.fastq.gz	Fastqsanger.gz

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Aurora-A interactome in MV4-11 cells

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Next generation sequencing identifies small RNA expression in FLT3-ITD+ AML (MV4-11) cell lines

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Next Generation Sequencing Facilitates Quantitative Analysis of MV4-11 cell line Transcriptomes

Project description:Purpose: The goals of this study are to compare side population and main population cells transcriptome profiling (RNA-seq) . Methods: We sorted side population and main population cells by staining Hoechst 33342 and using Flow Cytometer (Beckman Coulter, Moflo XDP). 200,000 cells per sample were analysed in RNA-Seq experiments.The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. qRT–PCR validation was performed using TaqMan and SYBR Green assays

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Project description:Enhanced homing and uncontrolled expansion are the characteristics of AML cells. We identified a transcription factor FEV (fifth Ewing variant) as the regulator. To assess the regulation mechanisms, FEV expression was knocked down in AML cell line MV4-11 cells using shRNA. MV4-11 cells transduced with non-silencing control (NC), FEV shRNA (shFEV) were cultured for 3 days and sorted by FACS. The cells were collected and total RNA was isolated by the Trizol kit (Takara Bio, Otsu, Japan) according to manufacturer’s protocol. High-throughput RNA sequencing (RNA-seq) was performed by Illumina HiSeq 2500 (Illumina, San Diego, CA) at CapitalBio Corporation (Beijing, China). Differentially expressed genes were analysed between the two groups.

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