Figure 4.

Treatment of PC12 cells with BFA results in inhibition of palmitoylation and membrane association of newly synthesized SNAP-25. PC12 cells were labeled with [³⁵S]methionine or [³H]palmitate for 1 h in the presence or absence of 10 μg/ml BFA. (A) Radiolabeled SNAP-25 was immunoprecipitated and analyzed by SDS-PAGE and fluorography. (B) [³⁵S]methionine-labeled cells were homogenized and fractionated into particulate (P100) and soluble (S100) fractions. The presence of SNAP-25 in each of the fractions as well as in the total cell lysate (TCL) was detected by immunoprecipitation followed by SDS-PAGE and fluorography.