Figure 8.

Intracellular mCD14–EGFP is rapidly exocytosed. Adherent U373–CD14–EGFP (□) and U373–EGFP (▵) cells cultured in 96-well plates were incubated with LPS–sCD14 complexes (40 ng/ml LPS) at 37°C for the times indicated. At 0 min, no LPS was added. At the end of the incubation, total fluorescence associated with the cells was measured in a fluorescent plate reader. Trypan blue (200 μg/ml) was added to quench extracellular fluorescence, and the remaining intracellular fluorescence was measured. Intracellular fluorescence was calculated as a percentage of total fluorescence in each well. Results are expressed as the means of triplicate wells ± SEM and are from an experiment performed three times with the same result. *Direct comparison using Student's t test shows that these values are significantly different from the value of cells incubated 0 min with LPS (P < 0.003).