Protocol for analyzing invadopodia formation and gelatin degradation

• Published in: STAR protocols 2024-12, Vol.5 (4), p.103399, Article 103399
• Main Authors: Meng, Weijian, Huang, Qingwen, Zhang, Rui, Shen, Jie, Xie, Daxing, Xu, Cong, Liu, Liang
• Format: Article
• Language: English
• Subjects: Animals; Cancer; Cell Line, Tumor; Cell Membrane; Extracellular Matrix - metabolism; Fluorescent Antibody Technique - methods; Gelatin - chemistry; Gelatin - metabolism; Humans; Microscopy; Neoplasm Invasiveness; Podosomes - metabolism; Protocol
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2024.103399

Invadopodia are actin-rich protrusions on the tumor cell membrane that degrade the extracellular matrix and play a crucial role in tumor cell invasion and metastasis. Here, we present a protocol to examine invadopodia’s ability to form and degrade the extracellular matrix during tumor invasion and metastasis. We detail the procedure for using immunofluorescence staining to indirectly detect invadopodia formation and assess their extracellular matrix degradation capability via the gelatin degradation assay. For complete details on the use and execution of this protocol, please refer to Huang et al.1 [Display omitted] •Detailed instructions for detecting invadopodia formation•A procedure for gelatin degradation assays and quantified by the software ImageJ•Assessment of invadopodia activity using the gelatin degradation assay•Cortactin and F-actin co-localization can be used to identify invadopodia formation Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Invadopodia are actin-rich protrusions on the tumor cell membrane that degrade the extracellular matrix and play a crucial role in tumor cell invasion and metastasis. Here, we present a protocol to examine invadopodia’s ability to form and degrade the extracellular matrix during tumor invasion and metastasis. We detail the procedure for using immunofluorescence staining to indirectly detect invadopodia formation and assess their extracellular matrix degradation capability via the gelatin degradation assay.