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The STAT3-Binding Long Noncoding RNA lnc-DC Controls Human Dendritic Cell Differentiation

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摘要 : Long noncoding RNAs (lncRNAs) play important roles in diverse biological processes; however, few have been identified that regulate immune cell differentiation and function. Here, we identified lnc-DC, which was exclusively expressed in human conventional dendritic cells (DCs).
Abstract: Long noncoding RNAs (lncRNAs) play important roles in diverse BioLogical processes; however, few have been identified that regulate immune cell differentiation and function. Here, we identified lnc-DC, which was exclusively expressed in human conventional dendritic cells (DCs). Knockdown of lnc-DC impaired DC differentiation from human monocytes in vitro and from mouse bone marrow cells in vivo and reduced capacity of DCs to stimulate T cell activation. lnc-DC mediated these effects by activating the transcription factor STAT3 (signal transducer and activator of transcription 3). lnc-DC bound directly to STAT3 in the cytoplasm, which promoted STAT3 phosphorylation on tyrosine-705 by preventing STAT3 binding to and dephosphorylation by SHP1. Our work identifies alncRNA that regulates DC differentiation and also broadens the known mechanisms of lncRNA action. The STAT3-Binding Long Noncoding RNA lnc-DC Controls Human Dendritic Cell Differentiation Fig. 4.lnc-DC directly binds STAT3 in cytoplasm to prevent Y705 dephosphorylation of STAT3 by SHP1. (A) RNA pull-down experiment with Mo-DC cytoplasmic extract. Specific bands were identified by MS (upper panel) or immunoblot of STAT3 (lower panel). (B) qPCR detection of the indicated RNAs retrieved by STAT3- or STAT1-specific antibody compared with immunoglobulin G (IgG) in the RIP assay within Mo-DC. (C) Colocalization analysis: RNA FISH assay of lnc-DC combined with immunofluorescence detection of STAT3 in Mo-DC. Scale bars, 10 mm. DAPI, 4′,6-diamidino-2-phenylindole. (D) Immunoblot detection of pSTAT3 in Mo-DC 5-day cultures with lentivirus-mediated lnc-DC RNAi or its control. (E) qPCR detection of lnc-DC retrieved by a STAT3-specific antibody in the RIP assay within Mo-DC pretreated with the STAT3 inhibitor S3I-201 for 30min. (F) Heat-map representation of the mean fold change in gene expression, as determined by transcriptome analysis of monocyte-derived cells with lentivirus-mediated lnc-DC knockdown or STAT3 inhibition (S3I-201) after 7 days DC culture (n = 2). Pearson correlation analysis: Pearson r = 0.63, P < 0.0001. DMSO, dimethyl sulfoxide. (G) Immunoblot of STAT3 Y705 phosphorylation after incubation of phosphorylated STAT3 with rhSHP1 in the presence or absence of lnc-DC RNA, sense (sen.) or antisense (ant.). *P < 0.05 and **P < 0.01 (two-tailed Student’s t test). 原文链接:http://www.ncbi.nlm.nih.gov/pubmed/24744378 作者:广州赛诚生物 点击:
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