Studying Cellular Focal Adhesion Parameters with Imaging and MATLAB Analysis
Journal
Bio-protocol
Journal Volume
13
Journal Issue
21
Start Page
e4867
ISSN
2331-8325
Date Issued
2023-11-05
Author(s)
Yu, Ling-Yea
Tseng, Ting-Jeng
Lin, Hsuan-Chao
Hsu, Chi-Lin
Lu, Ting-Xuan
Lin, Yu-Chiao
Tseng, Miranda
Abstract
Cell signaling is highly integrated for the process of various cell activities. Although previous studies have shown how individual genes contribute to cell migration, it remains unclear how the integration of these signaling pathways is involved in the modulation of cell migration. In our migration screen, we revealed that serine-threonine kinase 40 (STK40) and mitogen-activated protein kinase (MAPK) worked synergistically, and the suppression of both genes could further lead to suppression in cell migration. Furthermore, based on our analysis of cellular focal adhesion (FA) parameters using MATLAB analysis, we are able to find out the synergistic reduction of STK40 and MAPK that further abolished the increased FA by shSTK40. While FA identification in previous studies includes image analysis using manual selection, our protocol provides a semi-automatic manual selection of FAs using MATLAB. Here, we provide a method that can shorten the amount of time required for manual identification of FAs and increase the precision for discerning individual FAs for various analyses, such as FA numbers, area, and mean signals.
Subjects
Cell signaling
FA selection
Focal adhesion (FA)
Mean signal
Mitogen-activated protein kinase (MAPK)
Segmentation
Serine-threonine kinase 40 (STK40)
Thresholding
Type
journal article