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Blood, Vol. 110, Issue 7, 2545-2555, October 1, 2007 This Article Abstract Full Text Services Email this article to a friend Alert me to new issues of the journal Rights and Permissions Citing Articles Citing Articles via CrossRef JAM-C regulates unidirectional monocyte transendothelial migration in inflammation Blood Bradfield et al. 110: 2545 Supplemental materials for Bradfield et al, Vol. 110, Issue 7, 2545-2555 Files in this Data Supplement: Figure S1. Functional blocking of human JAM-B/-C by murine blocking antibody to JAM-C H33 (JPG, 34 KB) - (A) Chinese hamster ovary (CHO) cells transfected with JAM-B were labelled with a human JAM-C/Fc preclustered with PE-conjugated goat antihuman IgG-Fc. Biphasic expression of JAM-B on CHO cells with membrane bound human JAM-C Fc (black line) was not seen in a saline control (grey fill). (B) Inclusion of the murine mAb H33 led to a complete loss in fluorescence associated with bound JAM-C Fc. (C) This loss was not seen with the isotype control antibody 9B5. Figure S2. Electron micrographs of purified human monocytes prepared using isodensity centrifugation or negative selection (JPG, 83.2 KB) - (A) Monocytes purified by negative selection had no visible adherent platelets attached to membrane surfaces compared to monocytes prepared using isodensity centrifugation on NycoPrep (B), which contained several adherent platelets (indicated by black arrows) to a single monocyte. Figure S3. Capture and transmigration of human monocytes on activated HUVECs unaffected by a SolJAM-C (JPG, 24.1 KB) - A 5-minute bolus of monocytes was flowed over activated HUVECs at a shear stress of 0.05 Pa. Adherent monocytes were then counted and expressed as a mean of triplicate cell counts plus or minus SEM. No significant difference was observed with solJAM-C flag or a flag peptide control. Figure S4. Knockdown of JAM-C expression on HUVECs (JPG, 37.5 KB) - Histogram of JAM-C surface expression on HUVECs transfected with siRNAs against JAM-C (bold line) or a sham-transfected control (line). Immunostaining was performed with mAb H33 or isotype control mAb mcb-64 (dotted line). Video 1. Platelet deposition on luminal surfaces at the endothelial junction during human monocyte transmigration on HUVECs (WMV, 351 KB) - An adherent monocyte decorated with platelets undergoing transmigration simultaneously deposits adherent platelets (marked red) at the endothelial junction as it continues to migrate into the ablumen (green). Platelets remain adherent to the HUVEC luminal surface at the site of transmigration during the course of the flow assay (10 minutes). Video 2. Adherent human monocyte undergoing multiple transmigration events on cultured TNFα-activated HUVECs under flow (WMV, 1.41 MB) - The monocyte is marked in red or green when positioned on the luminal (phase grey) or abluminal (phase black) compartments, respectively, every fourth frame. This Article Abstract Full Text Services Email this article to a friend Alert me to new issues of the journal Rights and Permissions Citing Articles Citing Articles via CrossRef

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