Fig. 4.
Inhibition and kinetics of PR-176 formation in HLMs. (A) Inhibition of mEH activity by NSPA in HLMs. Pooled HLMs (0.5 mg/ml) were incubated with oprozomib (10 μM) or cis-SO (50 µM) in the presence of various concentrations of NSPA at 37°C for 15 minutes as indicated. The IC50 values were estimated using nonlinear regression data analysis. (B) Kinetics of PR-176 formation in male and female HLMs was conducted in 0.1 M potassium phosphate buffer (pH 7.4, 0.1 mg/ml BSA). Incubations were performed at 37°C for 30 minutes. Data were fitted into the Michaelis-Menten equation for calculation of the Km and Vmax values. Data represent mean ± S.D. from triplicate incubations.