Studying Cellular Focal Adhesion Parameters with Imaging and MATLAB Analysis
Ling-Yea Yu,
Ting-Jeng Tseng,
Hsuan-Chao Lin,
Chi-Lin Hsu,
Ting-Xuan Lu,
Yu-Chiao Lin,
Miranda Tseng,
Feng-Chiao Tsai
Affiliations
Ling-Yea Yu
Department of Pharmacology, National Taiwan University College of Medicine, Taipei, TaiwanDepartment of Internal Medicine, National Taiwan University Hospital, Taipei, Taiwan
Ting-Jeng Tseng
Department of Pharmacology, National Taiwan University College of Medicine, Taipei, Taiwan
Hsuan-Chao Lin
Department of Immunology, National Taiwan University College of Medicine, Taipei, Taiwan
Chi-Lin Hsu
Department of Pharmacology, National Taiwan University College of Medicine, Taipei, TaiwanDepartment of Internal Medicine, National Taiwan University Hospital, Taipei, Taiwan
Ting-Xuan Lu
Department of Pharmacology, National Taiwan University College of Medicine, Taipei, TaiwanDepartment of Pharmacology, National Taiwan University College of Medicine, Taipei, Taiwan
Yu-Chiao Lin
Department of Pharmacology, National Taiwan University College of Medicine, Taipei, Taiwan
Miranda Tseng
Department of Pharmacology, National Taiwan University College of Medicine, Taipei, Taiwan
Feng-Chiao Tsai
Department of Internal Medicine, National Taiwan University Hospital, Taipei, TaiwanDepartment of Pharmacology, National Taiwan University College of Medicine, Taipei, Taiwan
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 two-hit 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.
