Supplementary material for Clish et al. (1999) Proc. Natl. Acad. Sci. USA 96(14), 8247-8252.

Fig. 7. Methyl ester hydrolysis of ATLa₂ to free acid in ex vivo mouse whole blood. ATLa₂ was incubated ex vivo in mouse whole blood (2.8 mM) for 0, 2, 5, 10, 15, or 180 min. The incubations were stopped by chilling the blood on ice for 1 min followed by centrifugation (800 × g) at 0°C. The plasma supernatant was stopped in 2 vol of ice-cold methanol. The samples were prepared for analysis by solid phase extraction, and the amount of free acid in the sample was quantitated by liquid chromatography tandem MS. Within 10-15 min, 100% of the ATLa₂ methyl ester is hydrolyzed to the free acid without loss of compound. Values represent mean ± SEM (n = 3).