|
| Status |
Public on Apr 22, 2013 |
| Title |
GMT-treated MEFs, biological rep5 |
| Sample type |
RNA |
| |
|
| Source name |
zeocin-selected iCMs
|
| Organism |
Mus musculus |
| Characteristics |
transcription factor treatment: GATA4, MEF2C, TBX5
|
| Treatment protocol |
MEFs were plated into poly-L-lysine-coated 6 well dishes at 75,000 cells/well on Day -2, with 500 μL (≥ 2.5x105 IFU) each of rtTA lentivirus (FUdeltaGW-rtTA, Addgene plasmid 19780) and TroponinT-GCaMP-Zeo reporter lentivirus per well with 2 mL MEF medium. The following day (Day -1), culture medium was replaced with 2 mL fresh MEF medium and cells were transduced with 500 μL (≥ 2.5x105 IFU) of each tetO-transcription factor lentivirus. The next day (Day 0), medium was switched to 3 mL/well Reprogramming Medium consisting of AGM (Lonza CC-3186) without EGF, supplemented with 2 μg/mL doxycycline (Sigma).
|
| Growth protocol |
MEFs were grown in DMEM with 10% fetal bovine serum and 1X Glutamax.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Putative iCMs were enriched by selecting for zeocin resistance driven by the TroponinT-GCaMP5-Zeo reporter and collected in Trizol at Day 21 post-transduction (7 days after initiation of zeocin selection, 300 μg/mL). Non-transduced MEFs were cultured and collected at Day 21. Samples were collected in triplicate from independent biological replicates and total RNA was prepared by the University of Pennsylvania Molecular Profiling Facility.
|
| Label |
biotin
|
| Label protocol |
The labeling, hybridization, and scanning procedures were performed by the University of Pennsylvania Molecular Profiling facility
|
| |
|
| Hybridization protocol |
The labeling, hybridization, and scanning procedures were performed by the University of Pennsylvania Molecular Profiling facility
|
| Scan protocol |
The labeling, hybridization, and scanning procedures were performed by the University of Pennsylvania Molecular Profiling facility
|
| Description |
Gene expression data from iCMs generated from MEFs
GMT3
|
| Data processing |
Initial data analysis was performed using Affymetrix Microarray Suite 5.0 and further analyzed using PartekGS software.
|
| |
|
| Submission date |
Mar 19, 2013 |
| Last update date |
Apr 23, 2013 |
| Contact name |
Russell C Addis |
| E-mail(s) |
[email protected]
|
| Organization name |
University of Pennsylvania
|
| Department |
Cell and Developmental Biology
|
| Lab |
Gearhart Lab
|
| Street address |
3400 Civic Center Blvd, Bldg 421, SCTR 9-196
|
| City |
Philadelphia |
| State/province |
PA |
| ZIP/Postal code |
19104 |
| Country |
USA |
| |
|
| Platform ID |
GPL6246 |
| Series (1) |
| GSE45274 |
Optimization of Direct Fibroblast Reprogramming to Cardiomyocytes Using Calcium Activity as a Functional Measure of Success |
|
