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| Status |
Public on Oct 16, 2010 |
| Title |
Gene Expression Profile of 293T and 293T-Aire-expressing cells |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
To analyze the impact of Aire on gene expression profile in a model cell line, we used 293T cells and transfected them either with an Aire expression plasmid pCMV-Aire (where mAire is driven by CMV promoter) or with a control plasmid pCMV2B.
Total RNA was extracted 48 hours post transfection, processed and used for gene expression profiling by Affymetrix. The data demonstrate that Aire has a very broad impact, effecting (upregulating and downregulating) hundreds of differents genes, however these genes differ dramatically from its targets in medullary epithelial cells.
Keywords: transfection
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| Overall design |
293T cells were seeded on 10cm plates and were grown in DMEM containing 10% FBS under standard TC conditions. Next day, the cells were transfected either by pCMV-Aire (6ug/plate) or empty pCMV (6ug/plate) using Mirus reagent, according to manufacturer's instructions. Total RNA was extracted 48 hrs post transfection by Trizol protocol according to the manufacturer's instructions. 3 biological replicates were used for Aire transfection and 2 replicates for transfection with a control plasmid.
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| Contributor(s) |
Abramson J, Mathis D, Benoist C |
| Citation(s) |
20085707 |
| |
| Submission date |
Oct 15, 2010 |
| Last update date |
Mar 25, 2019 |
| Contact name |
CBDM Lab |
| E-mail(s) |
cbdm@hms.harvard.edu
|
| Phone |
617-432-7747
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| Organization name |
Harvard Medical School
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| Department |
Microbiology and Immunobiology
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| Lab |
CBDM
|
| Street address |
77 Avenue Louis Pasteur
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| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02215 |
| Country |
USA |
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| Platforms (1) |
| GPL570 |
[HG-U133_Plus_2] Affymetrix Human Genome U133 Plus 2.0 Array |
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| Samples (5)
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| Relations |
| BioProject |
PRJNA132421 |
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