|
| Status |
Public on Apr 25, 2010 |
| Title |
Nascent mRNA profiling of LPS-stimulated mouse macrophages |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
|
| Summary |
To identify transcriptionally regulated genes in primary mouse macrophages stimulated with LPS with high sensitivity, we isolated nascent RNA following metabolic labelling with 4-thiouridine during the last 35 min before cell harvest, as recently described (Dolken et al. 2008 RNA 14:1959-72). Microarray analyses of nascent RNA identified substantially more probe sets as up-regulated after 45 min of LPS stimulation than parallel analyses of total cellular RNA. In contrast, 4.5 h after stimulation, up-regulated genes in total and nascent RNA largely overlapped. This approach therefore allowed a much more sensitive detection of early changes in transcription, and the respective genes are likely to be direct targets of LPS-regulated transcription factors.
Keywords: Effect of LPS stimulation; comparison of changes in expression of total versus nascent RNA; timecourse
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| Overall design |
Two completely independent experiments were performed. Macrophages were stimulated with 100 ng/ml LPS for 45 or 270 minutes. Thiouridine pulsing was done in last 35 minutes before harvest. Total RNA was isolated. Nascent RNA was subsequently purified.
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| Contributor(s) |
Lang R, Frühauf K, Dölken L |
| Citation(s) |
20531401 |
| |
| Submission date |
Mar 08, 2010 |
| Last update date |
Mar 04, 2019 |
| Contact name |
Roland Lang |
| E-mail(s) |
[email protected]
|
| Organization name |
University Hospital Erlangen
|
| Street address |
Wasserturmstr. 3-5
|
| City |
Erlangen |
| ZIP/Postal code |
91054 |
| Country |
Germany |
| |
|
| Platforms (1) |
| GPL6246 |
[MoGene-1_0-st] Affymetrix Mouse Gene 1.0 ST Array [transcript (gene) version] |
|
| Samples (12)
|
| GSM518579 |
newly transcribed_270' post LPS_experiment 1 |
| GSM518580 |
newly transcribed_270' post LPS_experiment 2 |
| GSM518581 |
newly transcribed_45' post LPS_experiment 1 |
| GSM518582 |
newly transcribed_45' post LPS_experiment 2 |
| GSM518583 |
newly transcribed_not stimulated_experiment 1 |
| GSM518584 |
newly transcribed_not stimulated_experiment 2 |
| GSM518585 |
total RNA_270' post LPS_experiment 1 |
| GSM518586 |
total RNA_270' post LPS_experiment 2 |
| GSM518587 |
total RNA_45' post LPS_experiment 1 |
| GSM518588 |
total RNA_45' post LPS_experiment 2 |
| GSM518589 |
total RNA_not stimulated_experiment 1 |
| GSM518590 |
total RNA_not stimulated_experiment 2 |
|
| Relations |
| BioProject |
PRJNA124935 |
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