Supplemental Material

This article contains the following supporting material:

• Figure 1 - BFA-induced retrograde transport of β1,4 galactosyltransferase is delayed in COG-deficient patient fibroblasts.
  (A) Control and patient fibroblasts were treated with 5 μg/mL BFA for 12 min. and the localization of β1,4 galactosyltransferase was analyzed by immunofluorescence. Bar, 20μm. (B) Stained cells within a population (200-300) were classified into one of four typical staining patterns reflecting the BFA-induced collapse of the Golgi into the ER. The relative percentage of cells in a representative experiment within these classifications is shown. (C) The percentage of cells with an exclusive ER staining pattern was plotted as a function of time. The data represent the average of three independent experiments; errors bars indicate standard deviations. The symbols are as follows: control, closed circles; P1, closed squares; COG7-corrected P1, closed triangles.

• Figure 2 - Recovery of Golgi localization of β1,4 galactosyltransferase is not substantially altered in COG-deficient patient fibroblasts following BFA washout.
  (A) Control and patient fibroblasts were treated with 0.25 μg/mL BFA for 60 min. followed by washout of the drug as described in the Methods section. The localization of β1,4 galactosyltransferase was followed by immunofluorescence. Bar, 20μm. (B) Stained cells within a population (200-300) were classified into one of four typical staining patterns reflecting the recovery of Golgi localization. The relative percentage of cells in a representative experiment within these classifications is shown. ^a Cells with single, concentrated remnant-like structures were counted as exhibiting full ER localization (#1r; see Methods).

• Figure 3 - Maturation of the GPI-linked protein CD59 is not affected in P1 fibroblasts.
  Control and patient fibroblasts were labeled with ³⁵S-cysteine and CD59 immunoprecipitated as described under the Methods section. The position of molecular weight markers is shown.