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Contributor Information

  • Name Simon Cook
  • Institute Babraham Institute

Tool Details

  • Tool name: HM3 Cell Line
  • Alternate names: Mitogen-activated protein kinase kinase kinase 3, MAPK/ERK kinase kinase 3, MEK kinase 3, MEKK 3
  • Tool type: Cell Lines
  • Tool sub-type: Continuous
  • Parental cell line: HEK 293
  • Organism: Human
  • Tissue: Kidney
  • Disease: Cancer; Inflammation
  • Growth properties: Adherent cell line
  • Conditional: Yes
  • Conditional description: Stable transfectant, conditional functionality: MEKK3 kinase activity and ERK1/2, JNK and p38 signalling pathways activated following 4-hydroxytamoxifen (4-HT) treatment of cells.
  • Description: HM3 cells are HEK 293 cells stably expressing conditional kinase MEKK3:ER* from the pBabePuro plasmid. MEKK3:ER* consists of the isolated kinase domain of MEKK3 fused in-frame to a modified form of the hormone binding domain of the estrogen receptor (hbER*) that can be de-repressed by 4-hydroxytamoxifen (4-HT) but not ?-estradiol. In this case the * refers to a point mutation that ablates estradiol binding but allows 4-HT binding. Activation of MEKK3:ER* leads to the strong activation of the JNK and p38 pathways and a weaker activation of ERK1/2. This cell line can be used to study the role and factors impacting on the JNK, p38 and ERK1/2 signalling pathways such as gene expression, cell proliferation, cell cycle arrest and cell death.. The cells are puromycin resistant. Conditional kinase activation of MEKK3:ER* can be induced with 100nM 4-HT.
  • Research area: Cancer; Apoptosis and autophagy; Cell biology; Cell signaling and signal transduction; Immunology
  • Production details: HEK 293 cells were transfected with the pBabePuro plasmid expressing â?‚ˆ?‚†MEKK3:ER*. Stable transfectants were selected using puromycin and ring cloning.
  • Cellosaurus ID: CVCL_0306

  • For Research Use Only

Target Details

  • Target: MEKK3

Application Details

Handling

  • Format: Frozen
  • Growth medium: Phenol red-free Dulbecco's modified Eagle's medium (DMEM) high glucose version, 2 mM L-glutamine, 10% fetal bovine serum (FBS), 2 ?g/ml puromycin.
  • Shipping conditions: Dry ice

Documentation

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References

  • •  Gilley et al. 2009. Cell Signal. 21(6):969-77. PMID: 19249353.
  • •  ERK1/2, but not ERK5, is necessary and sufficient for phosphorylation and activation of c-Fos.
  • •  Boughan et al. 2006. J Biol Chem. 281(17):11637-48. PMID: 16513653.
  • •  Nucleotide-binding oligomerization domain-1 and epidermal growth factor receptor: critical regulators of beta-defensins during Helicobacter pylori infection.
  • •  Todd et al. 2004. Oncogene. 23(19):3284-95. PMID: 14981547.
  • •  ERK1/2 and p38 cooperate to induce a p21CIP1-dependent G1 cell cycle arrest.