|
| Status |
Public on Oct 10, 2018 |
| Title |
Heart_E9.5_Hdac3 Mutant vs WT_rep4 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
E9.5 Mutant Hearts 4
|
| Organism |
Mus musculus |
| Characteristics |
tissue: Embryonic hearts
developmental stage: E9.5
genotype: Nkx2.5Cre; Hdac3f/f mutants
background strain: mixed CD1/B6/129
|
| Treatment protocol |
Nkx2.5Cre; Hdac3f/+ mice were crossed to Hdac3f/f, embryos dissected at 9.5dpc, hearts microdissected on ice, we obtained 4 independent replicates of Nkx2.5Cre; Hdac3f/f mutants and littermate controls.
|
| Growth protocol |
Mice were maintained on mixed CD1/B6/129 genetic backgrounds separated by 4-8 generations of interbreeding from pure parental strains. Littermate embryos were analyzed in all experiments unless otherwise noted. Institutional Animal Care and Use Committee approved all animal protocols.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
100ng of total RNA were primed with 7 µl of 100 µM T7 DNA primer at 65°C for 10 min, then reversed transcribed at 40°C for 2 h in the presence of 1.2ul of Affinity Script BlockMix (Agilent) and 100 µM each dATP, dTTP, dGTP
|
| Label |
Cy5
|
| Label protocol |
cDNA labeled at 40°C for 2 hours with 25 µM dCTP or 25 µM Cy3-label in the prescence of T7 RNA Polymerase (Agilent)
|
| |
|
| Channel 2 |
| Source name |
E9.5 Wild-type Hearts 4
|
| Organism |
Mus musculus |
| Characteristics |
tissue: Embryonic hearts
developmental stage: E9.5
genotype: Hdac3f/f controls
background strain: mixed CD1/B6/129
|
| Treatment protocol |
Nkx2.5Cre; Hdac3f/+ mice were crossed to Hdac3f/f, embryos dissected at 9.5dpc, hearts microdissected on ice, we obtained 4 independent replicates of Nkx2.5Cre; Hdac3f/f mutants and littermate controls.
|
| Growth protocol |
Mice were maintained on mixed CD1/B6/129 genetic backgrounds separated by 4-8 generations of interbreeding from pure parental strains. Littermate embryos were analyzed in all experiments unless otherwise noted. Institutional Animal Care and Use Committee approved all animal protocols.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
100ng of total RNA were primed with 7 µl of 100 µM T7 DNA primer at 65°C for 10 min, then reversed transcribed at 40°C for 2 h in the presence of 1.2ul of Affinity Script BlockMix (Agilent) and 100 µM each dATP, dTTP, dGTP
|
| Label |
Cy3
|
| Label protocol |
cDNA labeled at 40°C for 2 hours with 25 µM dCTP or 25 µM Cy3-label in the prescence of T7 RNA Polymerase (Agilent)
|
| |
|
| |
| Hybridization protocol |
Oligoarray control targets and hybridization buffer (Agilent In Situ Hybridization Kit Plus) were added, and samples were applied to microarrays enclosed in Agilent SureHyb-enabled hybridization chambers. After hybridization, slides were washed in Agilent GE wash buffers 1-3.
|
| Scan protocol |
Scanned on an Agilent G2565AA scanner. Images were quantified using Agilent Feature Extraction Software (version A.8.5.1.1).
|
| Description |
Biological replicate 4 of 4. Control embryonic stem cells, untreated, harvested after several passages.
|
| Data processing |
Agilent Feature Extraction Software (v 8.5.1.1) was used for background subtraction and LOWESS normalization.
|
| |
|
| Submission date |
Dec 12, 2014 |
| Last update date |
Oct 10, 2018 |
| Contact name |
Michael Patrick Morley |
| E-mail(s) |
[email protected]
|
| Phone |
215-898-2026
|
| Organization name |
Perelman School of Medicine at the University of Pennsylvania
|
| Department |
Penn Cardiovascular Institute
|
| Street address |
3400 Civic Center Blvd, Bldg 421
|
| City |
Philadelphia |
| State/province |
PA |
| ZIP/Postal code |
19104 |
| Country |
USA |
| |
|
| Platform ID |
GPL10333 |
| Series (2) |
| GSE64146 |
Histone deacetylase 3 deletion in the embryonic heart at day E9.5 |
| GSE64149 |
Genome-nuclear lamina interactions regulate progenitor cell behavior during cardiac development |
|
