Fig. 5. BICD N–terminus recruits dynein and dynactin. (A) COS-1 cells were transfected with the indicated HA-fusion constructs and used for immunoprecipitation with anti-HA antibodies. Immunoprecipitates were analyzed by western blotting, using antibodies against HA, p150Glued, dynamitin, dynein intermediate chain (DIC) and MAB1614 against kinesin heavy chain (KHC). Western blot analysis with anti-KHC SUK4 antibody produced a similar result (not shown). (B–I) HeLa cells were transfected with GFP–BICD-N–MTS (GFP signal shown in B, D, F and H) and stained for p150Glued, DIC and KHC (antibody SUK4, MAB1614 produced a similar result; data not shown). Cells expressing low (B) or high levels (D, F and H) of GFP–BICD-N–MTS are shown. Bar, 10 µm.