Figure 1

GGA overexpression leads to accumulation of swollen vacuolar structures that stain positive for Golgi markers. (A) HeLa cells were left untransfected or were transfected with vectors expressing Myc-tagged GGA1, GGA2 or GGA3, HA-tagged TSG-F or TSG-3', or GFP-Vps4EQ. For detection of endogenous GGA proteins (left panels), cells were stained using antibodies specific for GGA1, GGA2 or GGA3. For exogenous GGA or Tsg101 protein detection, cells were stained using anti-Myc or anti-HA antibodies, respectively. Cells were analyzed using the DeltaVision RT microscope. Images shown were obtained from one of 7-8 independent experiments. (B) HeLa cells were transfected with the GGA expression vectors, fixed 24 h posttransfection, and stained with antibodies recognizing TGN46 or GM130. Images were acquired using a DeltaVision RT microscope and colocalization was determined using the SoftWoRx colocalization module. Numbers represent average colocalization (R values) ± SD from 10-15 cells. Size bar represents 15 μm.