PKC is essential for neutrophil, but not macrophage functions required for host defense against fungal pathogens. The C-type lectin receptor dectin-1 and the integrin Mac-1 have key roles in controlling fungal infection. Here, we demonstrate that dectin-1- and Mac-1-induced activation of protein kinase C in neutrophils, independent of the Card9 adaptor, is required for reactive oxygen species production and for intracellular killing upon Candida albicans uptake. Protein kinase C was also required for zymosan-induced cytokine generation in neutrophils. In macrophages, protein kinase C deficiency prevented fungi-induced reactive oxygen species generation but had no effect on activation of TGF--activated kinase-1, an effector of Card9, or nuclear factor B activation, nor did it affect phagolysosomal maturation, autophagy, or intracellular C. albicans killing. In vivo, protein kinase C-deficient mice were highly susceptible to C. albicans and Aspergillus fumigatus infection, which was partially rescued with adoptively transferred wild-type neutrophils. Thus, protein kinase C activation downstream of dectin-1 and Mac-1 has an important role in neutrophil, but not macrophage, functions required for host defense against fungal pathogens.