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| Status |
Public on Jun 28, 2011 |
| Title |
The effects of Hmgb1 depletion on gene expression in human cells |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
High Mobility Group Box 1 (HMGB1) is a highly abundant and evolutionarily conserved non-histone chromatin component with the ability to bind and bend DNA. The protein is involved in fundamental nuclear processes including nucleosome sliding, transcription, replication, V(D)J recombination and DNA transposition. To assess the functional impact of HMGB1 on transcription we performed gene expression profile analyses of HeLa cells stably transfected with an anti-HMGB1 shRNA expressing plasmid or a control plasmid.
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| Overall design |
2 stable clones: control HeLa, Hmgb1-knocked down HeLa, 3 technical replicates. Hmgb1 knockdown HeLa cells were prepared by stable transfection with the plasmid Hmgb1shRNA-pSuperior.puro or, as a mock control, with the empty vector pSuperior.puro (Invitrogen). Transfection was made starting from 500 thousands (500K) or 5 milions (5mil) cells. Transfected cells were selected with puromycin and single resistant clones were picked, amplified and analyzed for HMGB1 expression by western blot. A clone containing less then 10% of the wt amount of HMGB1 was selected for further experiments. Total RNAs were extracted using Quiagen RNeasy kit and analyzed on the Illumina BeadsArray platform in 2 technical replica (2 arrays).
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| Contributor(s) |
Celona B, Agresti A, Bianchi ME, Rossetti G, Rossi RL, Pagani M |
| Citation(s) |
21738444 |
| Submission date |
Oct 23, 2009 |
| Last update date |
Aug 16, 2018 |
| Contact name |
Riccardo Lorenzo Rossi |
| E-mail(s) |
rossi@ingm.it
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| URL |
http://www.ingm.it
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| Organization name |
Fondazione INGM
|
| Department |
Integrative Biology
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| Lab |
Pagani
|
| Street address |
Via Trivulzio 10
|
| City |
Milano |
| State/province |
MI |
| ZIP/Postal code |
20149 |
| Country |
Italy |
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| Platforms (1) |
| GPL6947 |
Illumina HumanHT-12 V3.0 expression beadchip |
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| Samples (8)
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| Relations |
| BioProject |
PRJNA121677 |
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