miR-125b is an adhesion-regulated microRNA whose expression can suppress differentiation. (A) a panel of miRNAs were assayed by qPCR for expression at low (3K/cm2) or high (20K/cm2) seeding densities (B) miRNAs expression as a function of cell spreading; spread (continuous fibronectin) or round (32×32 um2 micropatterns). (C) miR-125b expression was induced by knockdown of beta5 integrin (D) miR-125b expression was strongly induced by cell suspension, suspended (Sus) versus attached (Att) cells (E) miR-125b levels as a function of differentiation, GM:growth media, OM: osteogenic media, AM: Adipogenic media, LD: low density, 3K/cm2, HD: high density, 20K/cm2. (F) miR-125b mimetic (miR mimic) suppressed adipogenic and osteogenic differentiation of hMSCs compared to a transfection control (siGLO) or to miR-125b knockdown cells (si-miR). Oil-red-O staining used to visualize lipid droplets, and Fast Blue staining to visualize alkaline phosphatase (ALP) activity following 1 week of differentiation.