EGF-induced MAPK Phosphorylation Assay

Cells were treated with 10× stock of an EGF (PeproTech) dilution series for 10 min. Cells were treated with varying concentrations of EGF for 10 min before the evaluation of phosphorylated MAPK amounts by immunofluorescence as previously described (73 (link)) using an Operetta high-content imaging system (PerkinElmer). Data are the average of replicate wells generated using the Columbus image data storage and analysis system (PerkinElmer).

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Shi T., Niepel M., McDermott J.E., Gao Y., Nicora C.D., Chrisler W.B., Markillie L.M., Petyuk V.A., Smith R.D., Rodland K.D., Sorger P.K., Qian W.J, & Wiley H.S. (2016). Conservation of protein abundance patterns reveals the regulatory architecture of the EGFR-MAPK pathway. Science signaling, 9(436), rs6.

Publication 2016

EGF Operetta high-content imaging system Columbus image data storage and analysis system

Cells Dilution Immunofluorescence Replicate

Corresponding Organization : Environmental Molecular Sciences Laboratory

Other organizations : Center for Systems Biology, Harvard University

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Variable analysis

independent variables

Concentration of EGF

dependent variables

Phosphorylated MAPK amounts

control variables

Treatment time (10 min)

controls

No positive or negative controls were explicitly mentioned.

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