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. 2014 Oct 20;124(11):5042–5056. doi: 10.1172/JCI71385

Figure 7. Exogenous MATN2 induces proinflammatory gene expression in macrophages via TLR signaling pathways.

Figure 7

(A) Expression of Il1b, Tnfa, and Nlrp3 in WT and Myd88 KO macrophages (mean ± SEM, qRT-PCR) following incubation with recombinant MATN2 (2 μg/ml) at indicated time points (n = 3 cultures per condition, 3 independent experiments). Results are shown relative to untreated controls (dotted line; ****P < 0.0001, **P < 0.01, *P < 0.05, 2-way ANOVA). (B) Immunoblot analysis of whole cell lysates for pro–IL-1β, pro–TNF-α, and NLRP3 of WT and MyD88 KO macrophages without or after 30, 60, and 240 minutes of MATN2 incubation. Data show 1 of 4 independent experiments with total ERK as loading control. (C) WT and MyD88 KO macrophage cultures (stained with Calcein AM) after 24-hour incubation with recombinant MATN2 (2 μg/ml). Note that only WT cells are activated following MATN2 addition. Scale bar: 25 μm. (D) Immunoblot analysis for phosphorylated NF-κB, p38, and ERK1/2 as well as total NF-κB, ERK1/2, p38, and actin of WT and Myd88 KO macrophage whole cell lysates untreated or after 10, 30, 60, and 240 minutes of MATN2 incubation. Data show 1 of 4 independent experiments.