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. 2015 Oct 16;10(10):e0140740. doi: 10.1371/journal.pone.0140740

Fig 10. Less FVIII is bound to secreted/anchored ULVWF strings from stimulated HUVECs compared to GMVECs.

Fig 10

HUVECs were stimulated with 100μM histamine for 2 min. Cells were then stained as described in the legend for Fig 9 with rabbit anti-VWF plus chicken anti-rabbit IgG AF-488 (green), and with mouse monoclonal anti-FVIII plus goat anti-mouse IgG AF-647 (red). Panels (A, C and E) show representative ULVWF strings with bound FVIII from merged images. The HUVEC-secreted/anchored ULVWF string in panel (A) has predominant VWF detection and further verifies the specificity of the VWF and FVIII antibodies and their distinct fluorescent signals. Dashed lines (that were moved away to not obscure the string image) indicate the measured portions of the string. Graphs (B from image A, D from image C, and F from image E) show the intensities from the 488-nm (VWF, green) and 647-nm (FVIII, red) channels measured along the ULVWF string (in pixels) from the corresponding merged image. In the 60× images (A and E) 100 pixels = 11.4 μm and in the 100× image (C) 200 pixels = 11.8 μm. The ratio of FVIII intensity/VWF intensity is shown for each ULVWF string. Images are representative of 4 experiments.