Cells contain CMV and SV40 viral DNA sequences

This cell line stably expresses the constitutively activated form of MEK1 and can be used in MEK-MAPK pathway studies

This cell line stably expresses constitutively activated form of MEK1(MEKDD), and can be used in MEK-MAPK pathway studies.xa0

This cell line was derived from metastatic tumor and can be used to study breast cancer metastasis.xa0

Glu-Glu tagged phosphorylation site MEK1 mutant (MEKDD), verified at ATCC.

constitutively activated form of MEK1, expressed

EpH4 1424.2 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).xa0 Cells were rederived from a metastatic tumor to the lung after selection with 1 mg/mL G418 in complete medium.

This cell line stably expresses constitutively activated form of MEK1(MEKDD, Asp218/Asp222 MEK1 phosphorylation site mutant), and can be used in MEK-MAPK pathway studies. This cell line was derived from metastatic tumor and can be used to studyxa0breast cancer metastasis.xa0

Activation of MEK1 is mediated through phosphorylation of Ser218 and Ser222 by members of the Raf family of kinases.xa0

This line related to four other cell lines: B-MEKDD 116 cell line (ATCC CRL-3069), EpH4 1424 cell line (ATCC CRL-3071), EpH4 1424.1 cell line (ATCC CRL-3209), and EpH4-Ev cell line (ATCC CRL-3063).

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbeccos 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.

Medium renewal: Every 2 to 3 days

Temperature: 37°C

Atmosphere: air, 95%; carbon dioxide (CO₂), 5%

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