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Ligand activity of TAG-related molecules and other endogenous lipids. Siglec5 reporter cells were stimulated with plate-coated lipids for 18 h. Induction of NFAT was analyzed by flow cytometry. A, C18 TAGs with saturated or unsaturated fatty acid chains. Chemical structures are shown in Fig. S3C. B, glycerol and C18 saturated or unsaturated fatty acids. C, C18:2 DAG and MAG. Chemical structures are shown in Fig. S3D. D, glycerophospholipids, such as phosphatidylcholine (PC), PE, phosphatidylserine (PS), phosphatidylinositol (PI), and PG. E, cardiolipin. F, <t>C16</t> <t>ceramide</t> and sphingomyelin. G, cholesterol and cholesterol ester (cholesteryl linoleate). H, 5-PAHSA and 9-PAHSA. Chemical structures are shown in Fig. S3E. I, recombinant Siglec5-Fc and Fc proteins were incubated with plate-coated cardiolipin or 5-PAHSA. Bound proteins were detected with anti-hIgG-HRP. Data are representative of at least three independent experiments and are presented as the mean ± S.D. (error bars). *, p < 0.05; **, p < 0.01.
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Ligand activity of TAG-related molecules and other endogenous lipids. Siglec5 reporter cells were stimulated with plate-coated lipids for 18 h. Induction of NFAT was analyzed by flow cytometry. A, C18 TAGs with saturated or unsaturated fatty acid chains. Chemical structures are shown in Fig. S3C. B, glycerol and C18 saturated or unsaturated fatty acids. C, C18:2 DAG and MAG. Chemical structures are shown in Fig. S3D. D, glycerophospholipids, such as phosphatidylcholine (PC), PE, phosphatidylserine (PS), phosphatidylinositol (PI), and PG. E, cardiolipin. F, <t>C16</t> <t>ceramide</t> and sphingomyelin. G, cholesterol and cholesterol ester (cholesteryl linoleate). H, 5-PAHSA and 9-PAHSA. Chemical structures are shown in Fig. S3E. I, recombinant Siglec5-Fc and Fc proteins were incubated with plate-coated cardiolipin or 5-PAHSA. Bound proteins were detected with anti-hIgG-HRP. Data are representative of at least three independent experiments and are presented as the mean ± S.D. (error bars). *, p < 0.05; **, p < 0.01.
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Ligand activity of TAG-related molecules and other endogenous lipids. Siglec5 reporter cells were stimulated with plate-coated lipids for 18 h. Induction of NFAT was analyzed by flow cytometry. A, C18 TAGs with saturated or unsaturated fatty acid chains. Chemical structures are shown in Fig. S3C. B, glycerol and C18 saturated or unsaturated fatty acids. C, C18:2 DAG and MAG. Chemical structures are shown in Fig. S3D. D, glycerophospholipids, such as phosphatidylcholine (PC), PE, phosphatidylserine (PS), phosphatidylinositol (PI), and PG. E, cardiolipin. F, <t>C16</t> <t>ceramide</t> and sphingomyelin. G, cholesterol and cholesterol ester (cholesteryl linoleate). H, 5-PAHSA and 9-PAHSA. Chemical structures are shown in Fig. S3E. I, recombinant Siglec5-Fc and Fc proteins were incubated with plate-coated cardiolipin or 5-PAHSA. Bound proteins were detected with anti-hIgG-HRP. Data are representative of at least three independent experiments and are presented as the mean ± S.D. (error bars). *, p < 0.05; **, p < 0.01.
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Ligand activity of TAG-related molecules and other endogenous lipids. Siglec5 reporter cells were stimulated with plate-coated lipids for 18 h. Induction of NFAT was analyzed by flow cytometry. A, C18 TAGs with saturated or unsaturated fatty acid chains. Chemical structures are shown in Fig. S3C. B, glycerol and C18 saturated or unsaturated fatty acids. C, C18:2 DAG and MAG. Chemical structures are shown in Fig. S3D. D, glycerophospholipids, such as phosphatidylcholine (PC), PE, phosphatidylserine (PS), phosphatidylinositol (PI), and PG. E, cardiolipin. F, C16 ceramide and sphingomyelin. G, cholesterol and cholesterol ester (cholesteryl linoleate). H, 5-PAHSA and 9-PAHSA. Chemical structures are shown in Fig. S3E. I, recombinant Siglec5-Fc and Fc proteins were incubated with plate-coated cardiolipin or 5-PAHSA. Bound proteins were detected with anti-hIgG-HRP. Data are representative of at least three independent experiments and are presented as the mean ± S.D. (error bars). *, p < 0.05; **, p < 0.01.

Journal: The Journal of Biological Chemistry

Article Title: Identification of lipophilic ligands of Siglec5 and -14 that modulate innate immune responses

doi: 10.1074/jbc.RA119.009835

Figure Lengend Snippet: Ligand activity of TAG-related molecules and other endogenous lipids. Siglec5 reporter cells were stimulated with plate-coated lipids for 18 h. Induction of NFAT was analyzed by flow cytometry. A, C18 TAGs with saturated or unsaturated fatty acid chains. Chemical structures are shown in Fig. S3C. B, glycerol and C18 saturated or unsaturated fatty acids. C, C18:2 DAG and MAG. Chemical structures are shown in Fig. S3D. D, glycerophospholipids, such as phosphatidylcholine (PC), PE, phosphatidylserine (PS), phosphatidylinositol (PI), and PG. E, cardiolipin. F, C16 ceramide and sphingomyelin. G, cholesterol and cholesterol ester (cholesteryl linoleate). H, 5-PAHSA and 9-PAHSA. Chemical structures are shown in Fig. S3E. I, recombinant Siglec5-Fc and Fc proteins were incubated with plate-coated cardiolipin or 5-PAHSA. Bound proteins were detected with anti-hIgG-HRP. Data are representative of at least three independent experiments and are presented as the mean ± S.D. (error bars). *, p < 0.05; **, p < 0.01.

Article Snippet: Tetradecane (172456), pentadecane (P3406), octadecane (O0652), eicosane (219274), docosane (134457), tetracosane (T8752), squalane (234311), cardiolipin (C1649), C16 ceramide (43799), sphingomyelin (S0756), cholesterol (C8667), cholesteryl linoleate (C0289), and linoleic acid (L1376) were purchased from Sigma-Aldrich.

Techniques: Activity Assay, Flow Cytometry, Recombinant, Incubation