Figure 5.

SEAP production by HEK-Blue TLR4 cells treated with Bv overnight. The following day, was measured at 630 nm. Because Bv is a poor TLR4 stimulant, the treatments were assayed at 10 min time intervals to assess stimulation or inhibition of innate NF-κB activation, as measured by dependent SEAP activity. This also demonstrated that Bv concentration has no direct effect on the measurement of SEAP activity. Otherwise, the cells used in these experiments were not stimulated with other TLR4 ligands and SEAP is measured as a baseline activity. Comparison of 10 min SEAP activity (QuantiBlue^TM conversion; OD 630 nm) to 60 min SEAP activity demonstrated that the baseline enzyme increased at a regular rate, regardless of Bv concentration. The increase in SEAP over the entire 60 min, and between the 20–30 min time intervals, were both found to be significant by ANOVA (p < 0.05), as shown by brackets and an asterisks (*). There were no significant differences (ANOVA) between media controls and the 20 or 40 μM Bv treated reporter cells at either the 20 or 30 min intervals (designated by arrows; p > 0.05).