Figure 8.

Kinetic evaluation of SEAP activity in culture supernatants derived from Hb-treated HEK hTLR4 cells. Cells were cultured at an initial density of 1 × 10⁵ cells/well and treated with 40 μM Hb (Triangles), 40 μM Hb plus 40 μM Bv (squares), or complete medium with FCS (diamonds). All Bv and Hb dilutions were made in complete medium plus FCS. SEAP was tested using QuantiBlue substrate (InvivoGen). The data are presented in kinetic format, wherein OD 630 nm was measured in reaction wells every 10 min. A kinetic analysis was performed to show any changes that might occur in both total SEAP activity and enzymatic rates (Hb slope = 0.040, R² = 0.98; Hb + Bv slope = 0.007, R² = 0.99; No Treatment slope = 0.0016, R² = 0.98). Bv treatment only was shown in Figure 5 to have little or no effect on baseline SEAP and is not reiterated here. Significance between lines is shown with a bracket and an asterisk (*) designating p < 0.05 and was based upon the ANOVA results between the regression lines for Hb treatment only and Hb + Bv treatment.