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Status |
Public on Dec 31, 2014 |
Title |
The gene expression changes associated with loss of KDM5 (lid) in Drosophila wing discs |
Organism |
Drosophila melanogaster |
Experiment type |
Expression profiling by array
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Summary |
To determine the gene expression changes associated with loss of KDM5, we carried out microarray analyses of dissected wing imaginal discs from wildtype (w1118) and kdm510424 homozygous mutant 3rd instar larvae (96hr AED). kdm510424 is >99% homozygous lethal and we are unable to detect KDM5 protein by Western blot, thus it is likely to be a strong loss-of-function allele. Hierarchical cluster analysis of our microarray data revealed that of the 18,500 transcripts represented on the array, 367 genes were up-regulated and 534 genes were down-regulated 1.5-fold or more in kdm5 mutants compared to wildtype (p<0.05). To highlight biological processes potentially regulated by KDM5, we determined which gene ontology (GO) terms were enriched within the differentially expressed genes. While a total of ten GO terms were significantly enriched (p<0.01), two functionally related and overlapping terms clearly had the largest number of genes: (1) response to stress and (2) oxidation reduction.
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Overall design |
About 60 dissected w1118 and kdm510424 wing imaginal discs (three replicates each group) were prepared respectively using TRIZOL (Invitrogen) followed by on-column digestion of DNA using the RNeasy Mini Kit (Qiagen). RNA quantity and quality were assessed with a Beckman Coulter DU 640 spectrophotometer and Agilent 2100 Bioanalyzer, following the manufacturer’s protocols. Biotinylated, single-stranded, cDNA was prepared from 100ng total intact RNA. cDNA was hybridized to Drosophila Genome 2.0 GeneChips (Affymetrix).
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Contributor(s) |
Liu X, Secombe J |
Citation missing |
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Submission date |
Jan 07, 2014 |
Last update date |
May 04, 2018 |
Contact name |
Julie Secombe |
E-mail(s) |
[email protected]
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Organization name |
Albert einstein college of medicne
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Department |
Genetics
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Lab |
Julie secombe
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Street address |
1300 Morris park ave. Ullmann 809
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City |
Bronx |
State/province |
United states |
ZIP/Postal code |
10461 |
Country |
USA |
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Platforms (1) |
GPL1322 |
[Drosophila_2] Affymetrix Drosophila Genome 2.0 Array |
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Samples (6)
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Relations |
BioProject |
PRJNA233622 |