VIFs display reduced filament alignment and increased MT association in the sparse peripheral vimentin cytoskeleton. (A and B) Top-down (A) and oblique (B) 3D renderings of VIFs (blue), MTs (dark gray), and plasma membrane (light gray) reconstructed from perinuclear (left) and peripheral (right) subvolumes. (C) Magnified inset of perinuclear reconstruction indicated by the white box in B. (D) Scatter plot indicating average peripheral VIF-to-VIF neighbor distances and relative orientations. The dashed line indicates 20° co-orientation threshold. (E) Distribution of average NND between VIFs and MTs in either perinuclear (blue) or peripheral (gray) VIF networks. (F) Rendering of peripheral VIFs color-coded by local alignment with MTs. VIF segments that are both in proximity to and aligned with a neighboring MT (<100 nm NND and <20° relative orientation) are coded red. All other nonaligned segments are coded white. MTs are shown in dark gray, and the plasma membrane beneath is indicated in light gray. Inset 1 indicates peripheral VIFs that locally couple to the MT over short segments. Inset 2 indicates a hotspot with extensive VIF-MT co-alignment.