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U-2 OS Gal4-p300 Cell Line

Info

Catalogue Number 151579
Antigen/Gene or Protein Targets Gal4-p300
Parental Line U-2 OS
Host Human
Disease Keywords Osteosarcoma
Model Transgenic
Relevance U2-OS cells expressing Gal4-p300. p300 is a transcriptional coactivator that functions as integrator of numerous signaling pathways and are utilized by many DNA binding proteins to facilitate transcriptional activation. p300 shares numerous conserved domains with CREB binding protein (CBP), which is also a transcriptional coactivator. These shared domains include a histone acetyl transferase (HAT) domain, a bromo domain, and three cysteine- and histidine-rich domains. CBP also interacts with the RNA polymerase II holoenzyme and p300/CBP both contain transcriptional activation domains that function independently of HAT activity.
Production Details U2-OS cells were co-transfected with both the Gal4p300CRD1 expression vector (zeocin selection marker, backbone: pcDNA4/TO) and the Gal4-EIB-luciferase reporter vector (neomycin selection marker, backbone: pCG4 without the CMV promoter) using Fugene 6 transfection reagent.24 hours after transfection, stable transfectants were selected using G418 3.0 mg /ml and Zeocin 3.0 mg/ml until massive cell death, and then grown under G418 0.5 mg /ml and Zeocin 0.5 mg/ml for further selection. Established single Zeocin-resistant and Neomycin-resistant clones were isolated and expanded to generate individual clonal cell lines. These cell lines were verified by testing each clone for induction of high luciferase activity after the depletion of SUMO-conjugating enzyme Ubc9.
Research Area Epigenetics & Nuclear Signalling, Gene Expression
Recommended Growing Conditions DMEM plus 10% Foetal Bovine Serum and 1% Penicillin-Streptomycin

References: 1 entry

Girdwood et al. 2003. Mol Cell. 11(4):1043-54. PMID: 12718889.


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