Figure 8.

ATP6V0C overexpression sequesters tetherin in LAMP-1– and CD63–positive compartments. 293T cells were transfected with HA-tagged tetherin expression vector alone or in combination with the FLAG-tagged ATP6V0C expression vector. Twenty-four h post-transfection, cells were fixed and stained with anti-HA (green), anti-FLAG (red), and 4′,6-diamidino-2-phenylindole (blue) (A); with anti-HA (green), anti-CD63 (red), and anti-FLAG (blue) (B); or with anti-HA (green), anti-LAMP-1 (red), and anti-FLAG (blue) antibodies (C). Images were acquired with a Delta-Vision RT deconvolution microscope. Colocalization between tetherin and ATP6V0C (A), tetherin and CD63 (B), and tetherin and LAMP-1 (C) was quantified by calculating the Pearson correlation coefficient (r values) ± S.D. from 20–30 cells. Scale bars, 15 μm.