Preferential motility of Kinesin-1 occurs along stable microtubules marked by acetylation and detyrosination of α-tubulin.

Single KHC(1-560)-3xmCit motors in live COS cells were imaged by TIRF microscopy. The cells were fixed and stained for retrospective immunofluorescence. SD Maps of the KHC(1-560)-3xmCit motility events were created from the time series (far left images) and compared to the fixed images of (A) total tubulin (middle panel) and acetylated tubulin (right panel), (B) total tubulin (middle panel) and detyrosinated tubulin (right panel), or (C) total tubulin (middle panel) and polyglutamylated tubulins (right panel). Scale bar, 4 µm. The far right panels indicate schematic representations of the overlap between the Kinesin-1 motility events in the SD Map (green lines) and the modified microtubules (red lines).