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Sample GSM580574 Query DataSets for GSM580574
Status Public on Oct 01, 2010
Title THP-1 Cells: Uninfected + CAM vs. C. burnetii NMII infected + CAM - bio rep1, tech rep1
Sample type RNA
 
Channel 1
Source name Total RNA was harvested from Uninfected THP-1 human monocytic leukemia cells treated with Chloramphenicol
Organism Homo sapiens
Characteristics treatment: Chloramphenicol
infection: uninfected
cell line: THP-1(TIB-202)
passage: < 8
cell type: Monocytic leukemia cells derived from peripheral blood
Treatment protocol 10μg/ml of Chloramphenicol was added 48hpi and total RNA was harvested at 72hpi
Growth protocol Non-adherent THP-1 human monocytic leukemia cells (TIB-202; ATCC) were propagated in RPMI 1640 medium (Gibco, Carlsbad, CA) supplemented with 1mM sodium pyruvate, and 10% fetal bovine serum (FBS) at 37°C in 5% CO2
Extracted molecule total RNA
Extraction protocol Total RNA extracted using Trizol following manufacturer's instructions. Total RNA (500 ng) from each sample was then amplified using an Epicentre® Biotechnologies (Madison, WI) TargetAmp™ 1-Round AminoallylaRNA Amplification Kit, yielding approximately 6-10μg of aminoallyl-aRNA (AA-aRNA).
Label Alexa Fluor® 555-GREEN
Label protocol Approximately 6-10μg of aminoallyl-aRNA (AA-aRNA) was used for labeling. Alexa Fluor® 555-GREEN (Invitrogen, Carslbad, CA) was used to label the uninfected AA-aRNA and CAM treated uninfected AA-aRNA, while Alexa Fluor® 647-RED (Invitrogen) was used to label the AAaRNA from the C. burnetii infected cells and also for the CAM treated AAaRNA from the C. burnetii infected cells.
 
Channel 2
Source name Total RNA was harvested from C.burnetii infected THP-1 human monocytic leukemia cells treated with Chloramphenicol at 48hpi for 24 hours
Organism Homo sapiens
Characteristics treatment: Chloramphenicol
infection: C. burnetii infected
cell line: THP-1(TIB-202)
passage: < 8
cell type: Monocytic leukemia cells derived from peripheral blood
Treatment protocol 10μg/ml of Chloramphenicol was added 48hpi and total RNA was harvested at 72hpi
Growth protocol Non-adherent THP-1 human monocytic leukemia cells (TIB-202; ATCC) were propagated in RPMI 1640 medium (Gibco, Carlsbad, CA) supplemented with 1mM sodium pyruvate, and 10% fetal bovine serum (FBS) at 37°C in 5% CO2.Purified C. burnetii NMII SCVs at a genome equivalent MOI of 15 were used to establish a synchronous infection
Extracted molecule total RNA
Extraction protocol Total RNA extracted using Trizol following manufacturer's instructions. Total RNA (500 ng) from each sample was then amplified using an Epicentre® Biotechnologies (Madison, WI) TargetAmp™ 1-Round AminoallylaRNA Amplification Kit, yielding approximately 6-10μg of aminoallyl-aRNA (AA-aRNA).
Label Alexa Fluor® 647-RED
Label protocol Approximately 6-10μg of aminoallyl-aRNA (AA-aRNA) was used for labeling. Alexa Fluor® 555-GREEN (Invitrogen, Carslbad, CA) was used to label the uninfected AA-aRNA and CAM treated uninfected AA-aRNA, while Alexa Fluor® 647-RED (Invitrogen) was used to label the AAaRNA from the C. burnetii infected cells and also for the CAM treated AAaRNA from the C. burnetii infected cells.
 
 
Hybridization protocol Labeled AA-aRNA (2μg) with a dye incorporation efficiency range of 18 -34 picomol/microgram, were mixed pair-wise and hybridized overnight to Human OneArray™ microarrays (Phalanx Biotech Group, Palo Alto, CA). Human OneArrays contain 32,050 oligonucleotides; 30968 human genome probes and 1082 experimental control probes formed as 60-mer sense-strand DNA elements. Arrays were hybridized, washed and dried rapidly according to the manufacturer’s instructions.
Scan protocol Signal intensity of the hybridized arrays were measured by ScanArray Express (PerkinElmer, Boston, MA, USA) and the images were processed using GenePix Pro version 4.0 (Axon, Union City, CA, USA)
Description Biological replicate 1 of 3 - Technical replicate 1.
Data processing The processed GenePix Pro 4.0 output was further analyzed using Loess-Global intensity dependent normalization through the GenePix Auto Processor (http://darwin.biochem.okstate.edu/gpap3/)
 
Submission date Aug 17, 2010
Last update date Aug 24, 2010
Contact name SAUGATA MAHAPATRA
E-mail(s) saugata.mahapatra@okstate.edu
Phone 405-744-2506
Fax 405 744 6790
URL http://microgenetics.okstate.edu/~eshaw/
Organization name OKLAHOMA STATE UNIVERSITY
Department MICROBIOLOGY AND MOLECULAR GENETICS
Lab OBLIGATE INTRCELLULAR PATHOGENESIS
Street address 307 LSE, OSU
City STILLWATER
State/province OK
ZIP/Postal code 74078
Country USA
 
Platform ID GPL6254
Series (1)
GSE23665 Human THP-1 Cells: Uninfected vs. Coxiella burnetii NMII infected compared to Uninfected with Chloramphenicol vs. Coxiella burnetii NMII infected with Chloramphenicol

Data table header descriptions
ID_REF
VALUE normalized log2 ratio (Alexa 647/Alexa 555) representing test/reference

Data table
ID_REF VALUE
PH_hs_0000002 -0.504625142
PH_hs_0000003 0.167188109
PH_hs_0000004 -0.910183625
PH_hs_0000005 -0.709079322
PH_hs_0000006 2.27042371
PH_hs_0000007 -0.175235112
PH_hs_0000008 0.174370921
PH_hs_0000009 1.107507659
PH_hs_0000010 -0.777822026
PH_hs_0000011 -0.974535267
PH_hs_0000012 -0.759484927
PH_hs_0000013 0.866588626
PH_hs_0000014 1.246267763
PH_hs_0000015 0.064900021
PH_hs_0000016 -0.217930371
PH_hs_0000017 0.175378217
PH_hs_0000018 0.503674537
PH_hs_0000019 -1.515849906
PH_hs_0000020 0.212444791
PH_hs_0000021 0.952994012

Total number of rows: 30968

Table truncated, full table size 797 Kbytes.




Supplementary file Size Download File type/resource
GSM580574_THP-1Un+CAM_-_In+CAM_B_rep1_T_rep1.gpr.gz 2.6 Mb (ftp)(http) GPR
Processed data included within Sample table

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