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. Author manuscript; available in PMC: 2018 Jan 1.
Published in final edited form as: J Immunol. 2016 Nov 16;198(1):318–334. doi: 10.4049/jimmunol.1502381

Fig. 7. VLA3 activation is significantly increased on Fn + β-glucan vs. Fn-coated surfaces and is mediated by CR3.

Fig. 7

(A) Human neutrophils assayed as in Fig. 2. After 30 min incubation, neutrophils were isolated and stained with a directly conjugated mAb for a VLA3. FACS histogram showing total VLA3 on Fn (red) and Fn + β-glucan (blue). (B) Human neutrophils assayed as in Fig. 2 in the presence of the non-function blocking VLA3 mAb ASC-1 or isotype control. After 30 min incubation, bright field images acquired at 20x; bar = 100μm. (C) Shown is a schematic demonstrating the loss of FRET between ORB membrane dye and FITC-conjugated VLA3 mAb upon the extension of the extracellular domain. (D) Human neutrophils assayed as in Fig. 2 in the presence or absence of CR3 blocking mAb (44abc) or isotype control. After 30 min incubation, neutrophils were isolated and stained with a directly conjugated mAb for a VLA3 (Asc-1). Neutrophils were then incubated with 0, 75, 200, or 400 nM ORB and then analyzed by FACS. Representative data are plotted as the fraction of donor mean fluorescence intensity in the absence of acceptor fluorophore (Fd) to that in the presence of acceptor fluorophore (Fda) on the y-axis (Fd/Fda) vs. ORB mean fluorescence on the x-axis.