Visualization of RAS/MAPK signaling in situ by the Proximity Ligation Assay (PLA)

Abstract

RAS/MAPK signaling responds to diverse extracellular cues and regulates a wide array of cellular processes. Given its biological importance, abnormalities in RAS/MAPK signaling cascade have been intimately implicated in numerous human diseases, including cancer. Herein, we describe a novel methodology to study activation of this pivotal signaling pathway. The Proximity Ligation Assay (PLA) is employed to monitor kinase–substrate interactions between MEK1 and HSF1, or MEK1 and ERK1 in situ.