FIGURE 5.

ARF1 inactivity causes Golgi disassembly induced by AG1478 on H4 cells. A, ARF1-GTP pulldown assay. H4 cells or Rat2 cells were transfected transiently with ARF1-GFP for 24 h, treated with DMSO (lanes 1 and 5), 14 μm AG1478 (lanes 2 and 6), 140 μm AG1478 (lanes 3 and 7), and 5 μm BFA (lanes 4 and 8) for 1 h before lysis. The upper panels show the amount of GTP bound ARF1-GTP, and the lower panels show the total amount of ARF1-GTP in cells. B, H4 cells were pretreated with control or 50 μm AlCl₃ and 30 mm NaF for 10 min as indicated and then treated with 14 μm AG1478 or 1 μm BFA as indicated together for 30 min. The cells were stained with an antibody against β-COP. Bar, 10 μm.