SA2_Nutlin
Mus musculus
Protocols: Primary MEFs were isolated from E12.5-13.5 mouse embryos and amplified under low oxygen conditions (3% O2). All cells were maintained at ambient oxygen in humidified cell culture incubator (37°C with 5% CO2) in high-glucose Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% fetal bovine serum (FBS, Sigma-Aldrich), 100 U/ml penicillin and 100 µg/ml streptomycin (Life technologies). MEFs were treated for 6.5 hours with either 10µM Nutlin3a in DMSO or DMSO (10µl) alone. Total RNAs were purified using the RNeasy kit and the Qiacube robot from Qiagen. The quality of RNAs and the p53-specific induction of p21 expression upon Nutlin3a treatment has been verified by Reverse Transcription - quantitative Polymerase Chain Reaction (RTqPCR) using the Superscript VILO cDNA synthesis kit from Invitrogen and the Absolute qPCR cybergreen mix from Thermo Scientific, respectively. qPCR was performed on the Lightcycler 480 from Roche. RNASeq libraries were prepared from total RNA using the TruSeq Stranded mRNA LT kit (Illumina) according to the manufacturer's instructions.
ENA-FIRST-PUBLIC
2020-08-25T04:06:23Z
ENA-LAST-UPDATE
2020-06-03T09:47:04Z
External Id
SAMEA6871716
INSDC center name
Institute of Molecular Oncology, Philipps-University Marburg
INSDC first public
2020-08-25T04:06:23Z
INSDC last update
2020-06-03T09:47:04Z
INSDC status
public
Submitter Id
E-MTAB-9171:SA2_Nutlin
age
12.5 to 13.5
broker name
ArrayExpress
cell type
mouse embryonic fibroblast cell
common name
house mouse
developmental stage
embryo stage
genotype
Trp S180A
organism part
mouse embryonic fibroblast cell
sample name
E-MTAB-9171:SA2_Nutlin
scientific_name
Mus musculus
sex
male
strain
B6.129S