Cis 12 linoleic acid cla10 12

Cis-9, trans-11 linoleic acid (CLA9,11), trans-10, cis-12 linoleic acid (CLA10,12), and linoleic acid (LA) (all from Cayman Chemicals, Cambridge, UK) were prepared as 1, 10, and 50 mM stock solutions in 100% ethanol. Before each experiment, the stock solutions were diluted in warm complete culture medium to yield final concentrations of 1, 10, and 50 μM. The corresponding control was a 0.1% ethanol solution diluted in complete medium. For the experiments, EA.hy926 cells were seeded in 96-well plates (for MTT assay, ELISA, and adhesion assay), 6-well plates (for gene expression, flow cytometry), or T25 flasks (for gas chromatography), cultured in complete medium and exposed to different FAs for 48 h. Based on the conditions optimised for studying inflammatory responses of cultured EA.hy926 cells (Figure 8), after the FA exposure period, cells were incubated with TNF-α (1 ng/mL; 20 units/mL) for 6 or 24 h, depending on the assay to be performed.

Cis-9, trans-11 linoleic acid (CLA9,11), trans-10, cis-12 linoleic acid (CLA10,12), and linoleic acid (LA) were purchased from Cayman Chemicals, Cambridge, UK. They were prepared as stock solutions (1, 10 and 50 mM) in 100% ethanol. Prior to each experiment, the stock solutions were diluted in warm complete culture medium to give a final FA concentration of 1, 10 and 50 μM; the final ethanol concentration was 0.1%. The corresponding control was a 0.1% ethanol solution diluted in complete medium. For the experiments, the EA.hy926 cells were seeded in 96-well plates (for cell viability, inflammatory mediator and adhesion assays), 6-well plates (for gene expression assessment) and T25 flasks (for FA analysis), cultured in complete medium and exposed to different FAs for 48 h.