|
| Status |
Public on Mar 12, 2015 |
| Title |
lung tumors from mice with knock out of SMAD4 and PTEN rep3 |
| Sample type |
RNA |
| |
|
| Source name |
CCSPi-Cre mice, Ptend/dSmad4d/d lung cancer
|
| Organism |
Mus musculus |
| Characteristics |
genotype: CCSPi-Cre Ptend/dSmad4d/d
tissue: lung tumor
disease state: Ptend/dSmad4d/d lung cancer
genetic background: mixed C57BL/6
|
| Growth protocol |
All animal experiments were performed in accordance with an IACUC approved protocol (Baylor College of Medicine, Houston, TX). Generation of CCSPiCre mice was carried out as previously described with some modifications. An iCre-SV40 poly A fragment was amplified from Cyp17iCre plasmid by PCR and subcloned immediately upstream to a Frt-flanked phosphoglycerol kinase neomycin-resistance cassette (PGK-Neo). The iCre-SV40 poly A-PGK-Neo fragment was again subcloned into a pGEM-4Z plasmid harboring a 5.3-kb mouse CCSP genomic sequence to generate the targeting plasmid. The targeting plasmid was then linearized and transfected into R1 ES cells by electroporation. Genomic DNA was isolated from Neomycin-resistant clones and digested with Nde I and Xho I and were screened by southern-blot using a 5’ external probe. Twenty two of ninety eight clones were targeted. Correctly targeted clones were microinjected into blastocysts from C57BL/6 mice. Chimeras were bred to C57BL/6 mice and germ line transmission of the iCre gene to F1 offspring were validated with PCR genotyping. The CCSPicre mice were crossed with Ptenf/f and Smad4f/f mice to generate Ptend/d,Smad4d/d and Smad4d/d Ptend/d mice.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Twelve-month-old adult mice were used for the microarray analysis. After sacrifice, lungs were harvested from wild type mice, and lung tumors were macrodissected from Ptend/dSmad4d/d mice. These lung tissues were stored at -80°C and total RNA was extracted using the TRIzol method (TRIzol reagent; Invitrogen, Carlsbad, CA) followed by purification using an RNeasy column (Qiagen, Valencia, CA). RNA quality was assessed by using the Agilent Model 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA).
|
| Label |
biotin
|
| Label protocol |
According to Affymetix protocol.
|
| |
|
| Hybridization protocol |
According to Affymetix protocol.
|
| Scan protocol |
According to Affymetix protocol.
|
| Description |
knockout
|
| Data processing |
Processed the CEL files using dChip (PM-MM model, Quantile normalization).
|
| |
|
| Submission date |
May 20, 2013 |
| Last update date |
Mar 12, 2015 |
| Contact name |
Chad Creighton |
| E-mail(s) |
creighto@bcm.tmc.edu
|
| Organization name |
Baylor College of Medicine
|
| Department |
Biostatistics, Ducan Cancer Center
|
| Street address |
One Baylor Plaza, Mail Stop: BCM305
|
| City |
Houston |
| State/province |
TX |
| ZIP/Postal code |
77030 |
| Country |
USA |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE47116 |
Ablation of Pten and Smad4 leads to metastatic lung carcinoma in a murine model. |
|
