Figure 7.

GCC185 is needed for MPR transport to the Golgi complex in vivo and in vitro. (A) In vivo assay for MPR transport in control, mock-transfected cells or cells depleted of 90% of endogenous GCC185 protein or 77% of Golgin-97. The amounts of GCC185 or Golgin-97 were determined by loading equal protein on a 7.5% SDS-PAGE gel followed by immunoblot, as shown in the insets at the top. Transport was for 120 min at 37°C. Complete (100%) transport represents 721 cpm for the GCC185 siRNA experiment and 348 cpm for the Golgin-97 experiment, normalized per milligram of protein in each sample. (B) In vitro transport of CD-MPRs in reactions containing no addition (control), GST alone or GST-GCC185 C-terminal 110 residues (94 μg/ml). Transport was at 37°C for 90 min; 100% transport corresponded to 763 cpm.