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Blood, Vol. 112, Issue 13, 5016-5025, December 15, 2008
Anti-inflammatory effects of an inflammatory chemokine: CCL2 inhibits lymphocyte homing by modulation of CCL21-triggered integrin-mediated adhesions
Blood Flaishon et al.
112: 5016
Supplemental materials for: Flaishon et al
Files in this Data Supplement:
- Figure S1. FACS analysis of CCR7 expression on T cells treated with medium or CCL2 (JPG, 40.1 KB)
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- Figure S2. Cytoskeleton rearrangement (JPG, 29.8 KB)
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(A–B) Naïve T cells were stimulated with ELC (0.4 mg/ml) (A) or CXCL12 (0.1 mg/ml) (B) for 15 seconds in the presence or absence of CCL2 (0.1 ng/ml), fixed and permeabilized, and their intracellular F-actin was stained with FITC-phalloidin. The change in polymerized actin was analyzed by FACS. Percent increase in actin polymerization was calculated as the polymerization of actin in the presence of chemokine stimulation/ polymerization of actin without chemokine ×100. The results presented are representative of three separate experiments.
- Figure S3 (JPG, 40.3 KB)
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Frequency and strength of adhesive tethers between human T cells (intact or CCL2 pre treated) interacting with VCAM-1 (A), each coated alone or coimmobilized with CCL21. Experiments were performed at a shear stress of 0.75 dyn/cm2 (on VCAM-1). (B) Human T cells (intact or CCL2 pre treated) were allowed to accumulate for 2 min at a shear stress of 0.75 dyn/cm2 (VCAM-1/CCL21) and their adhesion persistence (ability to resist detachment by application of constant high shear stress (5 dyn/cm2) was assessed at the indicated time points. Results are shown as percent of cells initially accumulated on the integrin ligands.
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