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Genome-wide siRNA screen of genes regulating the Lipopolysaccharide-induced TNF-alpha response in human macrophages - GSE83824

Purpose

The mammalian innate immune system senses many bacterial stimuli through the toll-like receptor (TLR) family. Activation of the TLR4 receptor by bacterial lipopolysaccharide (LPS) is the most widely studied TLR pathway due to its central role in host responses to gram-negative bacterial infection and its contribution to endotoxemia and sepsis. Here we describe a genome-wide siRNA screen to identify genes regulating the human macrophage TNF-? response to LPS. We include a secondary validation screen conducted with six independent siRNAs per gene to facilitate removal of off-target screen hits. We also provide microarray data from the same LPS-treated macrophage cells to facilitate downstream data analysis. These data provide a resource for analyzing gene function in the predominant pathway driving inflammatory cytokine expression in human macrophages.

Hypothesis

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Experimental Design

THP-1 cells were stimulated with 0 and 10 ng/ml of LPS separately for 4hrs. Three replicates for each condition.

Experimental Variables

Lipopolysaccharide exposure (2021 ICD-10-CM code* = Not Applicable)

Controls

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Methods

Signal data was extracted from the image files with the Gene Expression module (v. 1.9.0) of the GenomeStudio software (v. 2011.1) from Illumina, Inc. Signal intensities were converted to log 2 scale. Probes that didn't show detection p-value < 0.1 in at least 2 arrays were removed from the dataset. After filtering probes, quantile normalization was applied across all arrays.

Additional Information

https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE83824

Microarray
Illumina HumanHT-12 v4
6 Samples Loaded: 6
Human (Homo sapiens)
Macrophages , Monocytes
Lipopolysaccharide exposure
THP1 B5 cells were treated with either 0 or 10 ng/ml LPS for 4 hr. LPS was from Alexis Biochemicals, Salmonella minnesota R595 TLRgrade, ALX-581-008-L002.
THP1 cells were maintained in RPMI, 10%FBS, 10mM Hepes, and 2mM glutamine by standard procedures.
RNA was purified by Qiagen RNAeasy kit
Sample Set Spreadsheet
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Samples Preview
Sample ID !Sample_title cell type cell line incubation_time experimental replicate
GSM2219158 JSTHP1LPS0hRep1 Macrophage THP-1 00h batch 1
GSM2219159 JSTHP1LPS4hRep1 Macrophage THP-1 04h batch 1
GSM2219160 JSTHP1LPS0hRep2 Macrophage THP-1 00h batch 2
GSM2219161 JSTHP1LPS4hRep2 Macrophage THP-1 04h batch 2
GSM2219162 JSTHP1LPS00Rep3 Macrophage THP-1 00h batch 3
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File
Log2 Transformed
Raw Signal
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Sample ID !Sample Title Cell type Cell line Incubation Time Experimental replicate Lps Concentration
GSM2219158
JSTHP1LPS0hRep1
Macrophage
THP-1
00h
batch 1
Zero
GSM2219159
JSTHP1LPS4hRep1
Macrophage
THP-1
04h
batch 1
10ng/ml
GSM2219160
JSTHP1LPS0hRep2
Macrophage
THP-1
00h
batch 2
Zero
GSM2219161
JSTHP1LPS4hRep2
Macrophage
THP-1
04h
batch 2
10ng/ml
GSM2219162
JSTHP1LPS00Rep3
Macrophage
THP-1
00h
batch 3
Zero
GSM2219163
JSTHP1LPS04Rep3
Macrophage
THP-1
04h
batch 3
10ng/ml

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