Fig. 6. Disrupting lipid raft integrity inhibits LPS-mediated cellular activation.
CHO/CD14/TLR4 reporter cell line was either not stimulated (A) or stimulated
with 10 ng/ml of LPS in 5% HPS for 30 minutes either in the absence (B) or
presence (C) of 60 µg/ml nystatin. The induction of CD25 surface expression
was detected with FITC-CD25. Fluorescence was detected using a FACSCalibur
(Becton Dickinson), counting 10,000 cells per sample. The effect of
raft-disrupting drugs on TNF-
secretion was measured (D) by treated
monocytes isolated from the blood of healthy donors with 10 mM MCD (black
circles) or 60 µg/ml nystatin (white squares) prior to stimulation with 10
ng/ml LPS in 5% HPS. Control experiments with cells stimulated with LPS in the
absence of raft-disrupting drugs were also performed (eclipse). The effect of
different concentrations of nystatin on LPS-induced TNF- secretion was
also measured (E). TNF- secretion was measured using an ELISA. Each
data point represents a number of independent experiments.
