Synchronized temporal-spatial analysis via microscopy and phosphoproteomics (STAMP) of quiescence

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 phosphoproteomics) to track changes in cellular landscape occurring throughout G₀ transition and ciliogenesis. We synchronized ciliogenesis and G₀ transition in two cell models and combined microscopy with phosphoproteomics 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.