Figure 2.

GST pull-down assay. (A) Rabring7 was expressed as a GST-fusion protein and its binding to Rab4wt, Rab7wt, or Rab8wt was examined in a GST pull-down assay. The cell lysates from the EGFP-tagged Rab4wt-, Rab7wt-, or Rab8wt-expressing COS1 cells were incubated with GST-Rabring7 or GST alone immobilized on glutathione-beads. EGFP-Rabs bound to the beads were detected by Western blot with the anti-GFP antibody. The expression levels of EGFP-Rabs in the cell extracts were also monitored by Western blot with the anti-GFP antibody. (B) Rabring7 was expressed as a GST fusion protein and its binding to Rab7wt, Rab7Q67L, Rab7T22N, or Rab7N125I were examined by the same method described above.