Figure 4.

GRAF1-depletion induces hyper-tubulation of CD59-containing endosomes and Rab11-positive recycling endosomes. Cells grown on coverslips were either mock-treated (A,C,E) or treated with GRAF1-siRNA (B,D,F) for 72 h. (A,B and insets) Cells were incubated with mouse anti-CD59 antibody for 15 min. At 37°C, followed by a 1 min. acid stripping and fixation. Fixed cells were stained with Alex 568-goat anti mouse secondary antibody. After siRNA-treatment, cells were fixed and stained with either anti-Rab11 antibody (C,D) or anti-Rab5 antibody (E,F), followed by appropriate secondary antibodies. Bar, 10μm.