FIGURE 8.

Palmitoylation of SNAP25a and SNAP23 by DHHC2. EGFP-SNAP25a and EGFP-SNAP23 were co-transfected into HEK293T cells together with HA-DHHC2 or empty vector (control). Twenty-four hours after transfection, the cells were incubated in [³H]palmitic acid for 4 h, washed, and lysed. The resulting lysates were resolved by SDS-PAGE and transferred to nitrocellulose. The ³H signal was detected using an intensifier screen, and duplicate nitrocellulose membranes were probed with anti-GFP. The top panel shows representative blots, with the molecular weight standards indicated on the left side. The bottom panel shows averaged data ± S.E. (n = 6) for palmitoylation in the presence and absence of DHHC2. The palmitoylation level in the absence of DHHC co-expression was set as 1.