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 (G(0)). Many pathologies arise due to disruptions in tissue-specific G(0), yet little is known about the temporal-spatial mechanisms that establish G(0) 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 G(0) transition and ciliogenesis. We synchronized ciliogenesis and G(0) 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.