Mycobacterium tuberculosis Requires Cholesterol Oxidase to Disrupt TLR2 Signalling in Human Macrophages

Figure 8

IL-8 and IL-10 secretion and gene expression in Mtb-infected macrophages. (a, d) The macrophages incubated with an IRAK1/4 inhibitor or untreated were infected with the wild-type, ΔchoD, or ΔchoD-choD strains for 24 hours. The (a) IL-8 and (d) IL-10 secretion was assessed using a BD CBA kit. The data are presented as from 5 independent experiments. IRAK1/4 inhibitor vs. without IRAK1/4 inhibitor, ; ^#ΔchoD vs. wild-type or ΔchoD-choD, (all Mann-Whitney test). (b) The macrophages treated with siRNA (either nontargeting or TRAF6 targeting) or untreated were infected with the wild-type, ΔchoD, or ΔchoD-choD Mtb strains for 24 hours. The IL-8 secretion was assessed using a BD CBA kit. The data are presented as from 5 independent experiments. siTRAF6 vs. the control, ; ^#ΔchoD vs. wild type or ΔchoD-choD, ; ^##ΔchoD siTRAF6 vs. wild-type siTRAF6 or ΔchoD-choD siTRAF6, (all Mann-Whitney test). (c, e) Macrophages treated with siRNA (either nontargeting or TRAF6 targeting) or untreated were infected with the wild-type, ΔchoD, or ΔchoD-choD Mtb strains for 24 hours. The (c) IL-8 and (e) IL-10 mRNA levels were measured using qRT-PCR. The data are presented as the mean relative quantification (RQ); . RQ represents the fold change in the gene expression in the infected macrophages compared to the noninfected macrophages, calculated using the ABI 7900HT (RQ) Manager Software (v1.2) and DataAssist Software v3.01 (Thermo Fisher Scientific). siTRAF6 vs. the control, ; ^#ΔchoD vs. wild type or ΔchoD-choD, ; ^##ΔchoD siTRAF6 vs. wild-type siTRAF6 or ΔchoD-choD siTRAF6, (all -test). Control: Mtb-infected macrophages untreated with siRNA; siNT: Mtb-infected macrophages treated with nontargeting siRNA; siTRAF6: Mtb-infected macrophages with a silenced TRAF6 gene.

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