Fig. 4.

Phosphatidylcholine (PC) is not required for SchuS4 lipid-mediated inhibition of inflammation in mammalian cells. a Confirmation of absence of PC in SchuS4Δpcs mutant. Lipids from wild-type (SchuS4) and SchuS4Δpcs mutant were spotted onto silica plates at increasing concentrations, and developed with Chl: MeOH:H₂O (7: 3: 0.25). Bands were visualized using primuline and UV illumination. Crude lipids from SchuS4 or SchuS4Δpcs were added at the indicated concentrations to mouse bone marrow-derived macrophages (BMDM) (b) or human dendritic cells (hDC) (c) (3–4 wells/group), followed by the addition of R848 (5 ng/mL) or LPS (5 ng/mL), respectively. Mock-treated controls (–; Mock) were treated with EtOH diluted similarly to wells containing 1 µg/mL lipid. Supernatants were assessed for IL-12p40 by ELISA. Data were analyzed using one-way-ANOVA followed by Tukey's multiple-comparisons analysis. BMDM data represents 3 experiments of similar design. A–C indicate individual donors. Error bars represent SD.