Figure 4

PKCδ phosphorylation of LCN2 at T115 is reduced in neutrophils from Prkcd ^−/− mice. (A) An affinity-purified rabbit anti-phospho-LCN2 (T115) antibody detected PKCδ phosphorylation of recombinant LCN2 (P), but not unphosphorylated LCN2 (N). The antibody was not immunoreactive against the LCN2 T115A mutant before or after incubation with PKCδ. (B) Representative western blot showing less immunoreactivity with the phospho-LCN2 (T115) antibody in neutrophil lysates from Prkcd ^−/− mice than in lysates from WT littermates. (C) The ratio of P-LCN2 (T115) to LCN2 immunoreactivity was significantly reduced in neutrophils from Prkcd ^−/− compared with Prkcd ^+/+ mice (n = 3) (*p < 0.05, two-tailed t test). Error bars indicate SEM.