Fig. 5: Bacterial killing and phagocytosis studies in various cells.

a For bacterial killing in neutrophils, JE2, JE2Δnuc1 and JE2Δnuc1 complemented with plasmid-expressed nuc1 (JE2Δnuc1 pRB473-nuc1) were opsonized with 10% human pooled serum. Neutrophils were incubated with bacteria at an MOI = 0.1 for 1 h and bacterial survival was checked at 10 min, 20 min, 30 min, and 1 h after exposure to neutrophils. Fluorescence-labeled bacteria were incubated with neutrophils at an MOI of 1:2 and the phagocytosis index was calculated 1 h after incubation via FACS. b To investigate bacterial survival in RAW 264.7, RAW 264.7 was incubated with JE2, JE2Δnuc1, and JE2Δnuc1(pRB473-nuc1) at an MOI of 1:20 for 1.5 h. Extracellular and attached bacteria were removed by gentamicin and lysostaphin before lysing the cells. Neutrophils used in these experiments were obtained from 4 donors and the other experiments were performed at least three times; error bars indicate mean ± SEM. Statistical analyses were performed using one-way ANOVA with Dunnett’s posttest. Statistical significance: not significant, p > 0.05, *p < 0.05; **p < 0.01; and ***p < 0.001.