a knockdown cells was in an active state that can effectively engage downstream effectors essential for phagolysosome fusion. In addition, flow cytometric staining of phagosomes isolated from both control and p110a knockdown cells showed no significant differences in the levels of Rab7 or Vps41, or in the cellular or phagosomal levels of the t-SNARE Vti1p and the v-SNARE VAMP7. These SNAREs are part of the trans-SNARE complexes which were previously shown to be involved in mediating homoand heterotypic late endosome-lysosome fusion. The presence of normal levels of key HOPS components and SNAREs indicates that the blockage in delivery of lysosomal components to phagosomes in p110a knockdown cells is not due to a lack of membrane fusion machinery, but an upstream event regulated by the PI3K p110a. Discussion While there are clear lines of evidence pointing to a role for the class III PI3K, Vps34, in mediating phagosome maturation events, recent studies have suggested that other classes of these lipid kinases may contribute to this process and to endosomal trafficking in general. Regulation of vacuole maturation is thought to be influenced by the ligands and receptors engaged in phagocytosis, although only very recently have there been detailed studies examining the effects of specific ligand-receptor interactions on the evolving vacuole. PI3K p110a and Phagosome Maturation Previous work has indicated a need for class IA PI3Ks in phagocytosis per se of IgG-opsonized prey, and Vps34 in mediating phagosome maturation. However, there are few reports to date examining the role PI3Ks play in the phagosome maturation of prey taken up by other means. A recent article directly comparing actin formation as a result of beta-Mangostin chemical information uptake by FccR or CR3 receptors noted the formation of PIP3, and the contribution of class IA PI3Ks to actin polymerization on late stage phagosomes only in the case of 
CR3. These results illustrate differential requirements for PI3K dependent upon the specific phagocytic receptors that are engaged. In particular, this study highlighted the role of PIP3 in phagosome trafficking and the importance of class IA PI3K in development of the CR3 phagocytic vacuole. Our results indicate the involvement of a class IA PI3K in phagosome maturation, and suggest that the class IA isoform, p110a, plays a role in this process. In particular, we have shown that PI3K p110a is required for the delivery of lysosomal components to late stage phagosomes in the case of prey taken up by CR3 and non-specific phagocytic receptors. This advances the novel concept that multiple PI3K classes are involved in regulating phagosome PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/2221058 maturation, with a requirement for class IA PI3K 7 PI3K p110a and Phagosome Maturation activity in regulating the delivery of late endosomal/lysosomal components. At first glance, it might appear that our data were collected at relatively late time points, ranging from 2 to 6 h in some cases. In fact, this was not very unusual, as multiple studies that have used magnetic or latex beads have reported protracted phagosome maturation times, which may be due to the indestructible nature of the prey. Nonetheless, it is important to note that we observed significant differences in the acquisition of lysosomal LAMPs, b-galactosidase, and lysosome-associated fluorescent dextran in our p110a knockdown cells between 12 h post phagocytosis, and that these differences were only enhanced further at later time points. Our data indicates that phagosome
