Synchronized Temporal-spatial Analysis via Microscopy and Phospho-proteomics (STAMP) of Quiescence

ABSTRACT Coordinated cell cycle regulation is essential for homeostasis, with most cells in the body residing in quiescence (G0). Many pathologies arise due to disruptions in tissue-specific G0, yet little is known about the temporal-spatial mechanisms that establish G0 and its signaling hub, primary cilia. Mechanistic insight is limited by asynchronous model systems and failure to connect context-specific, transient mechanisms to function. To address this gap, we developed STAMP (Synchronized Temporal-spatial Analysis via Microscopy and Phospho-proteomics) to track changes in cellular landscape occurring throughout G0 transition and ciliogenesis. For the first time, we synchronized ciliogenesis and G0 transition in two cell models and combined microscopy with phospho-proteomics to order signals for further targeted analyses. We propose that STAMP is broadly applicable for studying temporal-spatial signaling in many biological contexts. The findings revealed through STAMP provide critical insight into healthy cellular functions often disrupted in pathologies, paving the way for targeted therapeutics. TEASER STAMP of signaling in quiescent cells unravels transient phosphorylations in cellular functions. Competing Interest Statement The authors have declared no competing interest. Footnotes We updated the text to include new data and discoveries that help both validate our approach and open up exciting new directions in the fields of quiescence and ciliary function. We updated the author list to include new members of the team that helped in generating these data.