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| Status |
Public on Jul 11, 2012 |
| Title |
Whole genome transcription profile of antigen receptor activated B cells expressing wildtype or Calmodulin (CaM) - resistant E12 |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
To elucidate the genome-wide role of Ca2+/ calmodulin inhibition of E2A in regulation of BCR activation, we performed DNA microarray analysis of activated splenic B cells expressing wildtype or CaM-resistant E12 mutant m8N47 with and without anti-IgM treatment for 3 hour.The expression of a remarkably large set of genes differed significantly.
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| Overall design |
Primary splenic B-lymphocytes were purified from mice heterozygous for the deletion of E2A gene and infected with retroviruses expressing either wildtype or mutant E12 after activation with CD40L plus IL-4 for 14 h. After 12 h incubation, the infection was repeated for 12 h, followed by incubation for a further 22 h post-infection in fresh complete medium with the stimulants to allow expression of GFP and E12. In addition to CD40L plus IL-4, the medium was supplemented with LPS (200 ng/ml) during retroviral infection incubations to improve infection efficiency. To study gene expression following B-cell receptor activation, anti-mouse IgM (2.5 μg/ml) was added for 3 hour. For DNA microarray analysis of infected B-lymphocytes, the infected cells were purified with a FACSAria cell sorter instrument (BD Biosciences) using their green fluorescence from expression of the GFP.
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| Contributor(s) |
Verma-Gaur J, Grundström T |
| Citation(s) |
22581853 |
| Submission date |
Feb 13, 2012 |
| Last update date |
Jan 16, 2019 |
| Contact name |
Thomas Grundström |
| E-mail(s) |
thomas.grundstrom@molbiol.umu.se
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| Phone |
+46 90 785 2531
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| Organization name |
Umeå University
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| Department |
Department of Molecular Biology
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| Lab |
TGN Lab
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| Street address |
Analysvägen 1
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| City |
Umeå |
| ZIP/Postal code |
901 87 |
| Country |
Sweden |
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| Platforms (1) |
| GPL6887 |
Illumina MouseWG-6 v2.0 expression beadchip |
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| Samples (12)
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| Relations |
| BioProject |
PRJNA152089 |
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