Figure 7. LPS‐induced cytokine production was reduced in Atx‐ko macrophages.

• A, B
  Peritoneal macrophages from Atx^{ΔΜΕ/ΔΜΕ} mice and Atx^+/+ littermates (A) or Raw264.7 cells (B) were stimulated with LPS (50 ng/ml, 8 h) in the presence/absence of the ATX inhibitor (50 μM, 30 min). The culture medium was collected for ELISA to measure the level of secreted cytokines. All assays were performed in triplicate, and data are shown as mean ± SEM.
• C–E
  Peritoneal macrophages were stimulated with LPS or vehicle (endotoxin‐free water), after which intracellular cytokine production was measured through FACS analysis. With the scatter dot plot, the gate was set so that the viable cells were selected to analyze the intensity of the fluorescence signal by flow cytometry. The overlay plot represents the cytometry data.
• F
  To induce sepsis via LPS injection (Shirey et al, 2013; Voss et al, 2016), age (9–10 weeks)‐ and sex‐matched Atx^{ΔΜΕ/ΔΜΕ} mice (n = 9) and Atx^+/+ littermates (n = 21) were injected with LPS (i.p. 25 mg/kg). Survival was monitored for up to 16 h, and analyzed by the Kaplan‐Meier method (Log‐rank P = 0.0004).
• G
  Blood samples were collected from Atx^{ΔΜΕ/ΔΜΕ} (n = 7) and Atx^+/+ (n = 6) mice 10 h after LPS injection, after which the serum TNFα protein level was measured. The data are shown as mean ± SEM.

Data information: The data (F and G) were analyzed with the results accumulated by 3 independent experiments. All other data are the representative from three independent experiments. **P < 0.01, ***P < 0.001 (Mann–Whitney U‐test).