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. 2003 Jan 20;160(2):201–212. doi: 10.1083/jcb.200207045

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Copyright © 2003, The Rockefeller University Press

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Figure 4. — Golgin-84 is a specific binding partner for rab1. (a) GST-tagged rab1, rab2, and rab6 were loaded with GDP or GTPγS and incubated with Golgi extract, and specifically eluted proteins were analyzed by Western blotting with antibodies to GM130, golgin-45, and golgin-84. (b) Full-length GM130 and golgin-84 lacking the transmembrane domain were tested for interaction in the yeast two-hybrid system with the following rab proteins carrying activating point mutations: rab1Q70L, rab2Q65L, rab5Q79L, rab6Q72L, and rab33bQ92L. Interactions between the indicated proteins results in growth on high selection medium. (c) Full-length and truncation mutants of golgin-84 were tested for interaction with rab1Q70L in the yeast two-hybrid system. The CT mutant corresponds to the ΔHead + CC construct described in Fig. 5 without the membrane anchor.