Sanchez-Rendon Julio Cesar, Hundsdorfer Lara, Bastounis Effie E
Interfaculty Institute of Microbiology and Infection Medicine, University of Tübingen, 72076 Tübingen, Baden-Württemberg, Germany; Cluster of Excellence EXC 2124 Controlling Microbes to Fight Infections, University of Tübingen, 72076 Tübingen, Baden-Württemberg, Germany.
Interfaculty Institute of Microbiology and Infection Medicine, University of Tübingen, 72076 Tübingen, Baden-Württemberg, Germany; Cluster of Excellence EXC 2124 Controlling Microbes to Fight Infections, University of Tübingen, 72076 Tübingen, Baden-Württemberg, Germany.
STAR Protoc. 2025 May 7;6(2):103816. doi: 10.1016/j.xpro.2025.103816.
Extracellular signal-regulated kinase (ERK) activity waves regulate critical processes like wound healing and bacterial pathogen dissemination by altering host cell motility and biomechanics. Here, we present a protocol for fluorescence resonance energy transfer (FRET) imaging of ERK activity in the nuclei of epithelial cells in a monolayer using an ERK biosensor. Moreover, we outline all image processing steps for the spatiotemporal quantification of single-cell ERK oscillations and wave propagation. Modifications, especially with respect to FRET imaging, may be necessary if different ERK biosensors are used. For complete details on the use and execution of this protocol, please refer to Hundsdorfer et al..
细胞外信号调节激酶（ERK）活性波通过改变宿主细胞的运动性和生物力学来调节诸如伤口愈合和细菌病原体传播等关键过程。在这里，我们展示了一种使用ERK生物传感器对单层上皮细胞核中ERK活性进行荧光共振能量转移（FRET）成像的方案。此外，我们概述了单细胞ERK振荡和波传播的时空定量的所有图像处理步骤。如果使用不同的ERK生物传感器，可能需要进行修改，特别是在FRET成像方面。有关此方案的使用和执行的完整详细信息，请参考Hundsdorfer等人的研究。
