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. 2017 May 25;13(5):e1006346. doi: 10.1371/journal.ppat.1006346

Fig 3. VZV rSgC interacts with the cell surface through a specific interaction with GAGs.

Fig 3

(A) Histograms showing the interaction of MHV-68 M3 (left panel), HSV-2 rSgG (middle panels) and VZV rSgC (right panels) with CHO-K1 cells (upper panels) or CHO-618 cells (lower panels). CHO-K1 cells contain GAGs whereas CHO-618 cells are devoid of GAGs. Surface-bound proteins were detected by flow cytometry using an anti His-tag antibody. Light grey histograms represent the signal obtained when no recombinant protein was used. Empty histograms represent the signal obtained with 100 ng of purified recombinant protein. (B) Graph showing the number of resonance units (R.U.) obtained when rSgC (alone or in the presence of increasing concentrations of heparin) was injected over an SA chip containing immobilised heparin. The maximum R.U., recorded at 90 seconds, is shown. The signal obtained with buffer alone was subtracted from the signal obtained with the samples containing rSgC. The ratios of rSgC:heparin used are indicated in the X axis. (C) Western blots showing binding of VZV rSgB (top blot), VZV rSgC (middle blot) or VZV rSgI (bottom blot) to heparin beads. Bound proteins were detected by Western blotting using an anti His-tag antibody. Binding was competed with increasing amounts of soluble heparin (0.1, 0.5, 1 and 2 mg). The input, corresponding to 1/10 of the starting material, is shown in the right lane. One representative experiment out of at least three independent experiments is shown in A-C. Abbreviations: Hep, heparin; Hep B, heparin beads; gp, glycoprotein; rSg, recombinant soluble glycoprotein; kDa, kiloDaltons.