Supplemental Material

This article contains the following supporting material:

• Supplemental Fig. 1 - Internalization of fluorescein dextran in the absence (control) or the presence of PEG-cholesterol (+PEG-Chol). Subconfluent and confluent CHO cells were incubated with 0.5 μM biotinylated PEG-cholesterol for 1 min (subconfluent) or 5 min (confluent) at 15 °C. Cells were then washed and incubated with 1 mg/ml (subconfluent) or 5 mg/ml (confluent) fluorescein dextran for 30 min at 37 °C. Bar, 10 μm.
• Supplemental Fig. 2 - Analysis of GDP and GTP bound to rab11. Subconfluent, confluent and MβCD treated confluent cells transfected with GFP-rab11 were labeled with ³²Porthophosphate for 3 h at 37 °C. Cell lysates were then divided to two aliquots. Anti-GFP rabbit IgG was added to one aliquot whereas preimmune rabbit IgG was added to the other. After immunoprecipitation, the nucleotide bound to rab11 were analyzed by TLC as described in Materials and Methods. The percentage of GTP bound rab11 were calculated and displayed in Table.