Fig. 6. Dynactin interacts with nucleoporins, vimentin and dynein during female pronuclear migration. (A-C) Western blots following immunoprecipitation (IP) of dynactin p150^Glued from either pronucleate-stage zygotes or unfertilized oocytes. Lanes on the blots are beads only (B), IP pellets (P), IP supernatants (S) and whole cell lysates from either zygotes (Z) or oocytes (O). (A) Five nucleoporins of molecular mass 270 kDa, 175 kDa, 62 kDa, 35 kDa and 18 kDa are detected in the zygote IP pellet and whole zygote lysate, as well as in the oocyte IP supernatant and whole oocyte lysate. Nucleoporins in the IP pellet show modest enrichment. (B) Vimentin, identified at 58 kDa, is enriched in the zygote IP pellet and detected in both zygote and oocyte IP supernatants, as well as in whole zygote and oocyte lysates. (C) Dynein (74 kDa) and dynactin (150 kDa) are enriched in zygote and oocyte IP pellets and detected in whole zygote and oocyte lysates. (D) Dynactin p150^Glued (green) and nucleoporin p62 (red) co-localize around the female (F) and male (M) pronuclei, and at cytoplasmic foci (merged channels appear yellow). (E) Dynactin p150^Glued (green) and vimentin (red) co-localize around the pronuclei (merged channels appear yellow), with vimentin showing additional branching in the region between and surrounding the pronuclei. Scale bar, 10 µm.