|
|
|
|
|
|
|
|
|||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | |||||
|
Fig. 4. D2D388-274 inhibits chemokine-induced monocyte-adhesion. Rapid adhesion of monocytes to fibrinogen-coated slides was assessed as described in Materials and Methods. Adherent cells were counted in four independent 0.2 mm2 fields by using video-imaging software. Triplicates were analyzed for each condition. The data are the mean±s.d. of triplicate evaluation (a total of twelve 0.2 mm2 fields). (A) Monocytes were stimulated with MCP-1 (2 µM), fMLP (50 µM) or the indicated amounts of D2D388-274 and MMK-1 (MMK) and allowed to adhere for 3 minutes. (B) Monocytes were left untreated (-) or pretreated for 15 minutes with D2D388-274 (1 µM), MMK (50 µM), D1 (1 µM), 
D2D3 (1 µM) or full-length suPAR (1 µM). The ability of the cells to adhere to fibrinogen-coated slides was determined following stimulation with MCP-1 (black bars) or fMLP (white bars). As control, D2D388-274 was added to the cells simultaneously with MCP-1 or fMLP stimulation (Ctr-MIX). Gray bar, basal adhesion. (C) For titration: pretreatment for 15 minutes with suPAR at concentrations as indicated. Gray bar, basal adhesion (CTR); black bars, MCP-1 (1 µM). (D) Monocytes were left untreated (-) or preincubated with either anti-FPRL-1 Ab 
FPRL-1) or control Ig (Ctr Ig) for 15 minutes on ice after which they were treated for an additional 15 minutes at 37°C with medium (-), 1 µM D2D388-274, or 50 µM MMK. The ability of the cells to adhere to fibrinogen-coated slides was then determined following stimulation with MCP-1 (black bars) or fMLP (white bars). Gray bar, basal adhesion. (E) Monocytes were incubated with medium (-), the Hck inhibitor PP2 (10 nM), the PI3K inhibitor LY (25 µM) and the p38 inhibitor SB (1 µM) for 30 minutes at 37°C. Thereafter, half of the cells were treated with D2D388-274 (1 µM) for an additional 15 minutes at 37°C. Following this, all cells were stimulated with MCP-1 (2 µM) and allowed to adhere to fibrinogen-coated slides for 3 minutes. Gray bar, basal adhesion.
|
