Fig. 4

Effects of specific inhibitors of phagocytosis-related kinases on phagocytosis and phosphorylation of these kinases in infected macrophages. RAW 264.7 cells were pretreated with PP2 (SFKs inhibitor), PF-431396 (Pyk2/FAK inhibitor, PF), LY294002 (PI3K inhibitor, LY), Akt1/2 kinase inhibitor (Akti) and cytochalasin D (F-actin inhibitor, CD) for 4 h, 1 h, 1 h, 1 h and 1 h, respectively. Phagocytosis (a) and the total amount as well as phosphorylation level of each signaling molecule (b) were then estimated by gentamycin protection assay and immunoblotting, respectively, 90 min after coincubation of the pretreated cells with the bacteria at MOI 10. YJ: YJ016, WT strain; HL: HL128, MD mutant. DMSO: 5% dimethyl sulfoxide, the solvent used as negative control. Data in a were analyzed by one-way ANOVA followed by Tukey’s test. n = 3. Bars labeled with ‘a’ are not significantly different from each other, but are significantly different from that labeled with ‘b’ (P < 0.05). Data in b are representative of three independent experiments