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Mus musculus

ABSTRACT: RNA-seq analysis of gene expression profile in Uhrf1 WT and KO germinal center B cells

PROVIDER: PRJNA397185 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR5898185_1.fastq.gz	Fastqsanger.gz
	SRR5898185_2.fastq.gz	Fastqsanger.gz
	SRR5898186_1.fastq.gz	Fastqsanger.gz
	SRR5898186_2.fastq.gz	Fastqsanger.gz
	SRR5898187_1.fastq.gz	Fastqsanger.gz

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RNA-seq analysis of gene expression profile in Uhrf1 WT and KO germinal center B cells

Project description:The goals of this study are to compare gene expression profiles of Uhrf1 WT and KO germinal center B cells and reveal the underlying mechanisms by which Uhrf1 regulates germinal center B cell responses. We found that upon Uhrf1 depeletion in germinal center B cells, most of the differentially expressed genes were upreguated.

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Project description:The goals of this study are to compare gene expression profiles of Setd2 WT and KO germinal center B cells and reveal the underlying mechanisms by which Setd2 regulates germinal center B cell responses. Further our study aims to compare gene expression profiles of Phf19 WT and KO germinal center B cells reveal the underlying mechanisms by which Phf19 regulates germinal center B cells responses. Finally, we combine the two profiles and investigate the relation shape betweeen Setd2 dependent regulation and Phf19 dependent regulation.

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RNA sequencing of WT and Uhrf1 KO iNKT cells

Project description:Deletion of Uhrf1 resulted in stage 1-specific defects during iNKT cell development. To investigate the molecular mechanism, we sorted WT and Uhrf1-KO stage 1 iNKT cells and performed RNA-seq. By comparing gene expression profile, we found metabolic defects in Uhrf1-KO stage 1 iNKT cells. The expression of CD71 (Tfrc), two subunits of CD98 (Slc3a2 and Slc7a5) and Glut3 (Slc2a3) was reduced in stage 1 iNKT cells. Besides, the downstream pathways of AKT-mTOR axis were significantly reduced. Collectively, our results suggest that Uhrf1 is required for iNKT cell development by regulating the Akt-mTOR signaling pathway. We first sorted WT and Uhrf1-KO stage 1 iNKT cells, extracted the mRNA and performed RNA-seq. We then analyzed the differentially expressed genes and performed KEGG pathway analysis. We used RT-PCR to verify the expression of the key nutrient related genes (Tfrc, Slc3a2, Slc7a5 and Slc2a3) and used flow cytometry to test the protein level of metabolic related molecules. Besides, we also analyzed the expression of genes of mTOR downstream pathways to demonstrate that Uhrf1 mediated AKt-mTOR axis regulates iNKT cell development.

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