p150Glued/Nip100-TAP was tagged at the endogenous locus in haploid wild-type and various mutant strains. A) Immunoblots of whole-cell lysates. The level of TAP-tagged p150Glued/Nip100 is similar in wild-type and mutant cells. Dynamitin/Jnm1 is absent from the jnm1 mutant as expected. Cdc3 is a loading control. B) Immunoblots of high-speed supernatants, the S100 fraction, of the whole-cell lysates. The amount of p150Glued/Nip100 is decreased in the jnm1 and yll049w mutants. C) Immunoblots of Nip100-TAP precipitates loaded so that each lane received similar amounts of Nip100. Dynamitin/Jnm1 and p24/Yll049w are important for the association of Arp1 and Arp10 with Nip100. Intensities of Cap2 and Act1 bands were not reproducibly above control (not shown). Arp10 and p24/Yll049w were tagged with 13-myc. Strain numbers: untagged, yJC5813; wild-type, yJC5814; yll04wΔ, yJC5475; jnm1Δ, yJC5817; arp1Δ, yJC5815 and arp10Δ, yJC5816.