SignificanceIn this study, we have determined spatiotemporal compartmentalization of mechanical properties and their associated cell activities within the HF-SC niche during quiescence and activation. The spatial compartmentalization of stiff and quiescent hair follicle stem cells and soft and primed HGs is distinct from the architecture of the epidermis, where the basal, proliferative SC layer is soft and the suprabasal, differentiated layer is stiff. We have identified miR-205, a versatile and potent regulator of the actin cytoskeleton, whose expression modulates actomyosin contractility and the dynamics of cell size changes. Together, this study has implicated the dynamic changes of cell size and subsequent cell cycle reentry as a mechanism to sense and respond to mechanical forces during tissue regeneration.