Figure 8.

H89, but not H8, inhibits Sec13 recruitment. HeLa cells grown to 50% confluence on 12-mm glass coverslips were permeabilized in 30 μg/ml digitonin, washed, and incubated on ice as described in MATERIALS AND METHODS. After 20 min, cells were incubated in 50 μl of cytosol (A), buffer (B), cytosol + 50 μm H89 (C), or cytosol + 120 μM H8 (D) for 30 min at 32°C. All incubations included an ATP-regenerating system + 0.5 mM GTPγS. (E) Quantitation of A, C, and D. See MATERIALS AND METHODS for method of quantitation.