Fig. 1. Localization of Rab22a and its mutant forms in BHK-21 cells. Rab22a or the Q64L and S19N mutants were expressed by transfection of constructs in pcDNA3.1, followed by immunofluorescent double stainings to visualize the expressed protein and compartmental markers by confocal microscopy analysis. Wild-type Rab22a (A,D,G) localized on the surface of large vacuole-appearing structures, as well as on the plasma membrane. The vacuolar elements were positive for EEA1 (B) but did not contain LAMP-1 (E) or Rab11 (H). The Q64L mutant (J) showed a similar distribution and colocalized with EEA1 (K); the S19N mutant (M) displayed in many cells a diffuse cytosolic appearing staining often with additional brightly stained structures of variable size and shape in the perinuclear region. This mutant did not affect the intracellular distribution of EEA1 or show specific colocalization with it (N). Overlays are shown in panels C,F,I,L and O. Bar=10 µm.

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