ECM-coated polyacrylamide hydrogels have been used extensively to study how cells sense and respond to the physical characteristics of their microenvironments because their tunable mechanical properties can mimic the physiological and pathological elasticity of tissues38,39. To investigate role of CD44 in stiffness-dependent motility, we generated FN-coated polyacrylamide hydrogels of elastic moduli ranging from 2–4 kPa (hereafter called “soft”) to 18–20 kPa (hereafter called “stiff”).
