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This line was derived from BALB/c mouse xenografts, initiated with EpH4 cells stably transfected with an expression vector containing Glu-Glu epitope-tagged phosphorylation site MEK1 mutant (MEKDD).  Cells were rederived from the primary mammary fat pad tumors after selection with 1 mg/mL G418 in complete medium.

stably expresses Glu-Glu tagged MEKDD, which is a constitutively activated form of MEK1 (Asp218/Asp222 MEK1 phosphorylation site mutant)

This line was derived from BALB/c mouse xenografts, initiated with EpH4 cells stably transfected with an expression vector containing Glu-Glu epitope-tagged phosphorylation site MEK1 mutant (MEKDD).  Cells were rederived from the primary mammary fat pad tumors after selection with 1 mg/mL G418 in complete medium.

Activation of MEK1 is mediated through phosphorylation of Ser218 and Ser222 by members of the Raf family of kinases. 

This cell line stably expresses constitutively activated form of MEK1 (MEKDD), and can be used in MEK-MAPK pathway studies and breast tumor in vivo studies. This cell line is tumorigenic, and can be used as a breast cancer in vivo model

This line is one of four related cell lines: B-MEKDD 116 cell line (ATCC CRL-3069), EpH4 1424.1 cell line (ATCC CRL-3209), EpH4 1424.2 cell line (ATCC CRL-3210) and EpH4-Ev cell line (ATCC CRL-3063).

The EpH 1424 cell line produces the constitutively activated MEK1 mutant MEKDD which has been tagged with Glu-Glu, verified at ATCC.

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
3. Add 1.0 to 2.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
   Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
5. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 4 x 10³ to 8 x 10³ viable cells/cm² is recommended.
6. Incubate cultures at 37°C. Subculture when the cell concentration is between 8 x 10⁴ to 1.5 x 10⁵ cells/cm².

Pinkas J, et al. MEK1 signaling mediates transformation and metastasis of EpH4 mammary epithelial cells independent of an epithelial to mesenchymal transition. Cancer Res. 62(16): 4781-90. PubMed: 12183438