Figure EV3. VLCFA‐GM3 species synergistically and selectively control human TLR4/MD‐2 activation.

• A
  Co‐stimulation of human monocytes by GM3 species plus various TLR ligands: LPS (0.13, 0.25 ng/ml), TLR4/MD‐2, Pam3CSK4 (0.25, 0.5 μg/ml), TLR1/2, Flagellin (10, 50 ng/ml), TLR5, R848 (0.25, 0.5 μg/ml), TLR7/8, MALP‐2 (0.5, 1.0 ng/ml), and TLR2/6. IL‐6 production in culture supernatant was quantified by ELISA.
• B–D
  Production of IL‐6 (B), TNF‐α (C), and IL‐12/23 p40 (D) in culture supernatant following co‐stimulation of monocytes by GM3 species plus LA506 (synthetic TLR4 ligand) (3, 10, 30 ng/ml).
• E
  Relative luciferase reporter activities of NF‐κB, AP‐1, and ISRE in response to LPS (+, 5 ng/ml; ++, 1 μg/ml), sCD14 (1 μg/ml), GM3 22:0 (5 μM), and their combinations. Relative luciferase activity of control condition was defined as 1 for every reporter gene.

Data information: Data shown are mean ± SD (n = 3, A–D; n = 4, E) analyzed by Tukey's multiple comparison test (A) or by two‐tailed unpaired t‐test (B–D). *P < 0.05 and **P < 0.01 for comparisons between indicated groups.