Peptidoglycan-Like Components in Z-100, Extracted from Mycobacterium tuberculosis Strain Aoyama B, Increase IL-12p40 via NOD2

Figure 5

NOD2-mediated effect of Z-100 on IL-12p40. (a) IL-12p40 concentration in the BMDM culture supernatant in the presence of Z-100 (100 μg/mL) and a Syk inhibitor (100 nM). Data show the ; per group, pooled from 3 experiments. Statistical analysis was performed using Student’s -test (vs. control; NS: not significant). (b) IL-12p40 concentration in the human CD14⁺ cell culture supernatant in the presence of Z-100 (100 μg/mL) and a Syk inhibitor (100 nM). Data show the ; per group, pooled from 2 experiments. Statistical analysis was performed using Student’s -test (vs. control; NS: not significant). (c) Schematic of the knockdown experimental design. Human CD14⁺ cells were cultured with GM-CSF for 6 days before being seeded onto siRNA custom plates (NOD2 siRNA#1, NOD2siRNA#2, and negative control siRNA) and incubated for 48 hours. After 48 hours, stimulants were added to the medium, and the cells were cultured for 24 hours. (d) IL-12p40 concentration in the culture supernatant of human CD14⁺ cells with siRNAs. The cells were stimulated with Z-100 (10 or 100 μg/mL), PG (2 μg/mL), or MDP (10 or 30 ng/mL). Data show the ; per group, pooled from 3 experiments. ^## vs. saline treatment in the negative control siRNA transfection group (Steel’s multiple comparisons test). , , and vs. negative control siRNA transfection in each stimulation condition (PG and MDP 10 ng/mL group; Dunnett’s multiple comparisons test, other groups; Steel’s multiple comparisons test).

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