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. Author manuscript; available in PMC: 2010 May 24.
Published in final edited form as: Circ Res. 2008 Mar 6;102(8):950–958. doi: 10.1161/CIRCRESAHA.107.170779

Figure 6.

Figure 6

S1P modulates NFκB activation in macrophages. A, NFκB reporter activation. RAW264.7 macrophages were transfected with a NFκB-luciferase reporter plasmid. After 48 hours, cells were incubated in media alone (Ctrl), 10 ng/mL LPS (+LPS), or LPS +500 nmol/L S1P (+LPS+S1P) for 4 hours. Luciferase activity was then measured. *P<0.001 vs Ctrl, **P<0.001 vs LPS by ANOVA. B, Bone marrow–derived macrophages from C57BL/6J mice were incubated with 10 ng/mL LPS, 500 nmol/L S1P, or 1 μmol/L SEW2871 for 2 hours. Nuclear and cytosol extracts were collected and analyzed by SDS-PAGE for NFκB p65 and IκBα. A representative immunoblot is shown. Top, Nuclear extract probed with anti-NFκB p65 antibody. Bottom, Cytosolic extract probed with anti-IκBα antibody. C, Densitometry. Graphical representation of immunoblots in B normalized to histone H1 (nuclear) and tubulin (cytosol) from 3 independent experiments. *P<0.001 vs Ctrl, #P<0.003 vs LPS, **P<0.001 vs LPS by ANOVA.