Figure 3.

Calsyntenin-1 and Kinesin-1 interact in live cells. (A) HeLa cells were triple-transfected with EGFP-calsyntenin-1, HA-KLC1, and myc-KHC. Cell lysates were subjected to immunoprecipitation using the indicated antibodies. The tripartite complex was isolated with R85, anti-HA, and anti-c-myc antibodies, but not with nonspecific IgG. (B) Immunoprecipitation of calsyntenin-1 from the V1 membrane fraction of P7 mouse brains (see Supplementary Methods). Antibody R85 and R140 immunoprecipitates contained KLC1, KLC2, and KHC, but not JIP-1, SNAP-25, synaptophysin, and GAP-43. Nonspecific IgG did not precipitate any of these proteins. Note that longer exposure times were required to detect KLC and KHC in R140 immunoprecipitates (insets).