Supplemental Material

This article contains the following supporting material:

• Figure 1 - Furin localization is not disrupted upon GCC185 depletion. Antibodies were rabbit anti-GCC185 (left) and goat anti-furin (right). Bar indicates 20μm.

• Figure 2 - Localization of p115 is not significantly altered upon GCC185 depletion. Control and GCC185-depleted cells were labeled with rabbit anti-GCC185 (top row) and mouse anti-p115 (bottom row). The images were acquired on a deconvolution microscope and a total summation of the Z sections is shown. Approximate cell outlines are drawn in white.

• Figure 3 - Oligosaccharide maturation of the His- and Myc-tagged CD-MPR is normal in cells depleted of GCC185. Cells stably transfected with the CD-MPR were metabolically labeled and chased for the indicated times. CD-MPRs were recovered with Ni-NTA agarose beads, analyzed by SDS-PAGE and detected by autoradiography. Indicated at right is the mobility of a 50 kDa marker. The CD-MPR is a 31,000 MW polypeptide that is highly glycosylated; a shift to terminally glycosylated forms is seen in both samples when time zero samples are compared with later time points.