KBP increases the motility of KIF1Bα. A, extracts were prepared from HeLa cells that were either untransfected or transfected with KIF1Bα and KBP or KBPΔ251–281. The cells were cracked and cytosolic extracts separated by SDS-PAGE (20 μg of total protein) transferred onto nitrocellulose and blotted with KIF1B (top panel), KBP (midddle panel) and ubiquitous kinesin antibodies (bottom panel). B, uncoated or KIF1B antibody coated beads moving on MT sedimented on coverslips in flow cells were counted. Movements of beads were observed using VE-DIC in HeLa extracts as described in the Methods section. Each bar represents the number of beads counted during 5 minutes of observation on 10 planes in 3 independent experiments, where each plane was observed for 30 sec. C, distances traveled by beads in a particular extract were determined using RETRAC software. The total number of analysed beads (n) is indicated below each histogram.
Wozniak et al. BMC Cell Biology 2005 6:35 doi:10.1186/1471-2121-6-35