Both the cytoskeletal intermediate filaments (IFs) and cytoplasmic caveolae contribute to active pro- cesses such as cell migration, morphogenesis and vesicular trafficking, but the interplay between these two systems has remained elusive. Here, we find that vimentin and nestin IFs interact with caveolae central component caveolin-1 (CAV-1) and importantly, restrain the intracellular trafficking of CAV-1 positive vesicles by serving as a physical barrier. Consequently, CAV-1 vesicles show less density and mobility in vimentin IFs enriched region, which is a substrate stiffness independent process. Moreover, depletion of vimentin IFs releases the slow movement proportion of CAV-1 positive vesicles and thus increases their cytoplasmic dynamics, whereas the expression of caveolae-associated protein CAV-1, CAV-2 and cavin-1 were unaffected. Collectively, these results reveal a negative role of IFs in regulating the trafficking of intracellular CAV-1 vesicles in live cells. (C) 2018 Elsevier Inc. All rights reserved.