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| Status |
Public on Apr 01, 2018 |
| Title |
Transcriptional and Epigenomic profile of GSK126 or dox-mediated Ezh2 inhibition in KrasG12D/+;Trp53-/-;Ezh2i-GFP-2A-rTA;Luc lung tumors in vivo |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
We wanted to understand the consequences of GSK126-mediated Ezh2 inhibition in an orthotopic model of Kras-driven non-small cell lung cancer (NSCLC). We injected the NSCLC cells with above-mentioned genotype into Nude mice and treated them with GSK126 50mg/kg (daily) or vehicle. As additional control for Ezh2 specificity we treated one tumor with doxycycline that induces shRNA-mediated Ezh2 protein downregulation in those cells. Purified tumour cells were obtained by dissection and FACS sorting based of GFP expression.
This experiment contributes the genome-wide response of NSCLC cells to Ezh2 inhibition in vivo.
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| Overall design |
We generated mRNA profiles of tumor cells tail vein injected into the lungs of Nude mice by deep sequencing. After FACS purification, RNA extraction and Bioanalyzer analysis, we processed only samples with high quality cellular and RNA profiles. Overall, we compared 10-day GSK126 treated cells (n=4) and up to 30 days GSK126 treated cells (n=3) to Captisol-treated samples (vehicle, n=2), using Illumina Hiseq2000. FACS sorted cells from individual animals were obtained by GFP expression.
For H3K27ac and H2AK5ac profiling, we used KP primary tumors generated by injection of NSCLC into the tail vein of nude mice. Mice were sacrificed on the onset of shortness of breath and tissues were resuspended in ChIP lysis buffer.
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| Contributor(s) |
Gargiulo G, Serresi M, Cesaroni M, de Rink I, Company C |
| Citation(s) |
30487290 |
| |
| Submission date |
Sep 04, 2014 |
| Last update date |
Mar 25, 2019 |
| Contact name |
Gaetano Gargiulo |
| E-mail(s) |
gaetano.gargiulo@mdc-berlin.de
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| Organization name |
Max Delbrüch Center
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| Department |
Molecular Oncology
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| Street address |
Robert-Rössle-Straße 10
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| City |
Berlin |
| ZIP/Postal code |
13125 |
| Country |
Germany |
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| Platforms (2) |
| GPL13112 |
Illumina HiSeq 2000 (Mus musculus) |
| GPL19057 |
Illumina NextSeq 500 (Mus musculus) |
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| Samples (18)
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| Relations |
| BioProject |
PRJNA260292 |
| SRA |
SRP046252 |
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