Figure 6.

Endogenous and overexpressed GGA1 proteins localize to the perinuclear region in NRK cells. NRK cells were cultured, fixed, and permeabilized as described in MATERIALS AND METHODS. (A) The rabbit polyclonal GGA1 antiserum (R-79709) was used as the primary antibody at a dilution of 1:500 to increase the signal from endogenous protein. (B) Specific staining was prevented by previous incubation of the primary antibody with antigen. (C) Exposure of the cells to BFA (30 μM) for 1 min. (D) Staining very similar to that of endogenous protein was seen after the GGA1-GST fusion protein was introduced into cells by microinjection.