Fig. 3. MLK3 interacts with the N-terminal region of golgin-160. (A) EGFP-tagged golgin-160 deletion mutants containing N-terminal regions spanning amino acids 1-59 (lane 1), 1-139 (lane 2) or 96-393 (lane 3) were transiently expressed with wild-type MLK3 in CHO cells. Following GFP immunoprecipitation, protein complexes were resolved by SDS-PAGE and immunoblotted for MLK3 (top) and GFP (bottom). (Left) The MLK3 and EGFP-tagged golgin-160 proteins in the immunoprecipitations. (Right) The expression of MLK3 and EGFP-tagged golgin-160 proteins in the total cell lysates (10% of the input for the immunoprecipitations). Data represent three separate experiments. (B) MLK3 interacts with GST-tagged golgin-160(1-393). MLK3 or Akt-1 were transcribed and translated in vitro in the presence of ³⁵S-methionine. GST or GST/golgin-160(1-393) pre-bound to glutathione-agarose beads were incubated with the labeled proteins and bound material was resolved by SDS-PAGE and autoradiography. The input is 10% of the starting material added to each pull-down. These data are representative of three independent experiments.