Ekasit SonphoFrederick G. MannMichaella LevyEric J. RossCarlos Guerrero-HernándezLaurence FlorensAnita SarafViraj DoddihalPuey OunjaiAlejandro Sánchez AlvaradoThailand Ministry of EducationMahidol UniversityHoward Hughes Medical InstituteStowers Institute for Medical Research2022-08-042022-08-042021-08-01Molecular and Cellular Proteomics. Vol.20, (2021)15359484153594762-s2.0-85117948677https://repository.li.mahidol.ac.th/handle/20.500.14594/76073The extracellular matrix (ECM) is a three-dimensional network of macromolecules that provides a microenvironment capable of supporting and regulating cell functions. However, only a few research organisms are available for the systematic dissection of the composition and functions of the ECM, particularly during regeneration. We utilized the free-living flatworm Schmidtea mediterranea to develop an integrative approach consisting of decellularization, proteomics, and RNAi to characterize and investigate ECM functions during tissue homeostasis and regeneration. ECM-enriched samples were isolated from planarians, and their proteomes were characterized by LC–MS/MS. The functions of identified ECM components were interrogated using RNA interference. Using this approach, we found that heparan sulfate proteoglycan is essential for tissue regeneration. Our strategy provides an experimental approach for identifying both known and novel ECM components involved in regeneration.Mahidol UniversityBiochemistry, Genetics and Molecular BiologyChemistryArticleSCOPUS10.1016/J.MCPRO.2021.100137