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. 2018 Dec 3;9:2761. doi: 10.3389/fimmu.2018.02761

Figure 9.

Figure 9

NLRX1-TUFM is required for activation of MAPKs-AP-1 pathway and proinflammatory cytokine response to H. capsulatum. (A–C) Macrophages from WT and Nlrx1−/− mice were stimulated with or without (0 min or medium) H. capsulatum (MOI = 5). Cell lysates were collected at 15, 30, 60, and 120 min after stimulation and subjected to Western blotting for the analysis of the indicated proteins (A). Relative intensity of indicated protein normalized against the corresponding β-actin was shown in (B) (n = 3). Supernatants were harvested at 18 h after stimulation, and the concentrations of TNF and IL-6 in the supernatants were quantified by ELISA (n = 11) (C). (D,E) Macrophages from WT mice were transfected with control siRNA or siRNA against TUFM (50 nM) for 72 h. Cells were then stimulated with or without (0 min) H. capsulatum (MOI = 5). Cell lysates were collected at 30 and 60 min after stimulation and assessed by Western blotting for the analysis of the indicated proteins (D). Supernatants were harvested at 18 h after stimulation, and the concentrations of TNF and IL-6 in the supernatants were quantified by ELISA and are presented as the relative levels of TNF and IL-6 (n = 6) (E). Bars represent the mean ± SEM, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001 [2-tailed t-test (B,E); ANOVA with Bonferroni's multiple comparisons post-hoc test (C)].