Multiplexed monitoring of MAP Kinase signaling
In this video, mouse fibroblast cells (NIH3T3) express Kinase Translocation Reporters (KTRs) for the three MAP Kinases in three different colors: ERK KTR in green, JNK KTR in red and p38 KTR in blue. Kinase Translocation Reporters are engineered kinase substrates that change localization upon phosphorylation. Basically, when the fluorescence is nuclear the kinase is inactive and when the fluorescence is cytoplasmic the kinase is active. Cells are imaged every 12 minutes. About 6 hours after the start of the movie, we add anisomycin which induces translational stress and activates all three MAP kinases. After 2 hours a specific JNK inhibitor is added. By the end of the movie an inhibition of ERK fluctuations is observed.