FIGURE 4.

Disruption of the TGN regulates the recycling of HA-β1AR to the plasma membrane after removal of isoprotenerol. Cells were prelabeled with rabbit HA mAb at 4 °C, and incubated at 37 °C with ISO and vehicle or BFA for 0, 1, 2, and 4 h. After agonist and surface receptor-bound antibody were removed, cells were further incubated with vehicle or BFA for 0.5 h (B–D and F–H) or 1 h (E and I). A–I, fixed cells were incubated with goat anti-rabbit Alexa 594 for visualizing total surface-labeled HA-β1AR (A, red) or recycled HA-β1AR (B–I, red) prior to permeabilization, and then incubated with goat anti-rabbit Alexa 488 for visualizing endocytosed HA-β1AR (green) post-permeabilization. Results are representatives from 3 independent experiments. J, ELISA showing the quantitative results of experiments similar to D and H.