Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of human KG1 cells differentiated towards dendritic like cells (DLCs) using PMA and TNF-alpha. Gene expression was compared between undifferentiated KG1 cells and day 4 DLCs

ABSTRACT: KG1 cells were differentiated towards dendritic like cells (DLCs) using PMA and TNF-alpha. Gene expression was compared between undifferentiated KG1 cells and day 4 DLCs.

ORGANISM(S): Homo sapiens

SUBMITTER: Jonathan Keller

PROVIDER: E-MEXP-345 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Gene expression analysis of hematopoietic progenitor cells identifies Dlg7 as a potential stem cell gene.

Gudmundsson Kristbjorn Orri KO Thorsteinsson Leifur L Sigurjonsson Olafur E OE Keller Jonathan R JR Olafsson Karl K Egeland Torstein T Gudmundsson Sveinn S Rafnar Thorunn T

Stem cells (Dayton, Ohio) 20070222 6

Inducible hematopoietic stem/progenitor cell lines represent a model for studying genes involved in self-renewal and differentiation. Here, gene expression was studied in the inducible human CD34+ acute myelogenous leukemia cell line KG1 using oligonucleotide arrays and suppression subtractive cloning. Using this approach, we identified Dlg7, the homolog of the Drosophila Dlg1 tumor suppressor gene, as downregulated at the early stages of KG1 differentiation. Similarly, Dlg7 was expressed in nor ...[more]

PMID: 17322106

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Project description:Dendritic cells (DC) arise from a diverse group of hematopoietic progenitors and have marked phenotypic and functional heterogeneity. We have found previously that activation of protein kinase C beta 2 (PRKCB2) by cytokines or phorbol esters drives normal human CD34(+) hematopoietic progenitors and myeloid leukemic blasts (KG1, K562 cell lines, and primary patient blasts) to differentiate into DC, but the genetic program triggered by PRKCB2 activation that results in DC differentiation is only beginning to be characterized. Of the cPKC isoforms, only PRKCB2 was consistently activated by DC differentiation-inducing stimuli in normal and leukemic progenitors. To examine early changes in gene expression following PRKCB2 activation, we employed the following cell lines: (1) the CD34(+) human acute myeloid leukemia derived cell line KG1, which undergoes DC differentiation following phorbol ester treatment; (2) KG1a, a spontaneously arising differentiation-resistant daughter cell line of KG1 that has lost PRKCB2 expression; (3) clones established from KG1a that stably express exogenous PRKCB2-GFP fusion proteins and are once again able to undergo DC differentiation (KG1a-PRKCB2-GFP Clone E9 and Clone E11). We examined changes in gene expression in these cells following treatment with the phorbol ester PMA (phorbol 12-myristate 13-acetate) for 2 hours. Since KG1 and KG1a differ in PRKCB2 expression but have similar expression of the other protein kinase C isoforms, this protocol will allow for the identification of genes regulated by PRKCB2 activation. KG1, KG1a, E9, and E11 were cultured for 2 hours in normal media +/- PMA (10 ng/ml). RNA was then isolated using RNeasy mini-columns (Qiagen) following the standard protocol. Samples were then sent to Expression Analysis, who carried out quality control, sample processing/labeling, hybridization to Affymetrix GeneChip arrays (Human U133 2.0 Plus), imaging, and analysis according to their standard protocols. Gene expression profiles were generated for each cell line in the presence and absence of phorbol ester stimulation, for a total of 12 samples (E9 +/- PMA and E11 +/- PMA were run in duplicate).

Dataset Information

Transcription profiling of human KG1 cells differentiated towards dendritic like cells (DLCs) using PMA and TNF-alpha. Gene expression was compared between undifferentiated KG1 cells and day 4 DLCs

Publications

Gene expression analysis of hematopoietic progenitor cells identifies Dlg7 as a potential stem cell gene.

Similar Datasets

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