QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

Help viewing high resolution images Return to article

Fig. 2. D2D3_88-274 stimulation does not induce CCR2 heterologous desensitization. (A) MCP-1 binding to CCR2 was measured on human monocytes and CCR2-overexpressing CHO cells by the addition of 1 nM ¹²⁵I-MCP-1 (see Materials and Methods). The percentage of control specific binding is indicated in the figure. (B) Monocytes and THP-1 cells were loaded with 1 mM Fura-2 for 30 minutes at 37°C and then stimulated with 1 µM MCP-1 or 1 µM D2D3_88-274. Fluorescence output was measured at 340 nm in a fluorescence spectrophotometer. The results were confirmed in two different experiments. (C) THP-1 cells were pretreated with serum-free medium (med), 10 nM D2D3_88-274 (D2D3) or 10 nM uPA (uPA) for 30 minutes and then stimulated with 100 nM MCP-1 for the times indicated (x-axis). At the end of the stimulation, cells were fixed, permeabilized and stained with FITC-labeled phalloidin. The amount of bound phalloidin-FITC was evaluated by flow cytometry analysis and its increase in percent, compared to the content of F-actin detected in unstimulated cells, is shown in the figure (y-axis). The data in A and C represent the mean±s.d. of three experiments. Each experiment was performed in triplicate for each stimulus.

Return to article