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Clinical

Microarray analysis of IL-10 stimulated adherent peripheral blood mononuclear cells-GSE43700

Purpose

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Hypothesis

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Experimental Design

Peripheral blood monuclear cells derived through Ficoll-Hypaque from the blood of healthy human donors (n=4). Cells adhered to tissue culture treated 6-well plates for 2 hours in 1% Fetal Bovine Serum (FBS) RPMI and then stimulated by IL-10 (R&D Systems) 10ng/ml or media alone in 10% FBS RPMI for 24 hours at 37°C, 5% CO2. After stimulation, cells harvested and monocytes isolated through CD14 positive selection (Miltenyi Biotec). RNA from purified monocytes extracted by Trizol and purified by Qiagen RNeasy Kit. RNA probe and microarray performed by UCLA Clinical Microarray Core using Ambion labeling kit and Affymetrix Human U133 Plus 2.0 array.

Experimental Variables

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Controls

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Methods

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Additional Information
  • Teles RM, Graeber TG, Krutzik SR, Montoya D et al. Type I interferon suppresses type II interferon-triggered human anti-mycobacterial responses.Science 2013 Mar 22;339(6126):1448-53. PMID: 23449998
  • Microarray
    Affymetrix HG-U133_Plus_2
    8 Samples Loaded: 8
    Human (Homo sapiens)
    Monocytes
    Healthy
    Sample Set Spreadsheet
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    Samples Preview
    Sample ID Donor ID !Sample_title cell type stimulated with
    GSM1068613 1 A6_media Peripheral blood monuclear cells media alone for 24hr
    GSM1068614 1 A7_IL-10 Peripheral blood monuclear cells IL-10 (10ng/ml) for 24hr
    GSM1068615 2 B6_media Peripheral blood monuclear cells media alone for 24hr
    GSM1068616 2 B7_IL-10 Peripheral blood monuclear cells IL-10 (10ng/ml) for 24hr
    GSM1068617 3 C6_media Peripheral blood monuclear cells media alone for 24hr
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    Raw Signal
    Raw Signal
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    Sample ID Donor ID !Sample Title Cell type Stimulated with
    GSM1068613
    1
    A6_media
    Peripheral blood monuclear cells
    media alone for 24hr
    GSM1068614
    1
    A7_IL-10
    Peripheral blood monuclear cells
    IL-10 (10ng/ml) for 24hr
    GSM1068615
    2
    B6_media
    Peripheral blood monuclear cells
    media alone for 24hr
    GSM1068616
    2
    B7_IL-10
    Peripheral blood monuclear cells
    IL-10 (10ng/ml) for 24hr
    GSM1068617
    3
    C6_media
    Peripheral blood monuclear cells
    media alone for 24hr
    GSM1068618
    3
    C7_IL-10
    Peripheral blood monuclear cells
    IL-10 (10ng/ml) for 24hr
    GSM1068619
    4
    D6_media
    Peripheral blood monuclear cells
    media alone for 24hr
    GSM1068620
    4
    D7_IL-10
    Peripheral blood monuclear cells
    IL-10 (10ng/ml) for 24hr

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