Figure 6.

PKCε phosphorylates NSF in vivo. A, An affinity-purified rabbit anti-phospho-NSF (S460) antibody was generated that detects recombinant NSF phosphorylated by PKCε (P), but not unphosphorylated NSF (N). The antibody was not immunoreactive against the NSF S460A mutant before or after incubation with PKCε in a kinase reaction. B, Representative Western blot showing that the anti-phospho-NSF (S460) antibody detected less immunoreactivity in brain lysates from Prkce^−/− mice than in lysates from wild-type littermates. C, The ratio of P-NSF (S460) to NSF immunoreactivity was reduced in samples from Prkce^−/− (n = 4) compared with Prkce^+/+ (n = 4) mice (*p < 0.05, two-tailed t test). Error bars indicate SEM.