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. 2016 Aug 5;11(8):e0160696. doi: 10.1371/journal.pone.0160696

Fig 4. Authentic RGD- and divalent cation-dependent binding of FMDV empty capsids to truncated, FLAG-tagged bovine αvβ6.

Fig 4

(A) Sandwich ELISA for the detection of FMDV EC. Wells were coated with cell culture supernatant from mock-transfected cells (Mock) or cells co-transfected with expression plasmids for the αv and β6-FLAG subunits (αvβ6) and incubated with FMDV A22ecRGD (RGD) or A22ecKGA (KGA). FMDV capsid proteins were detected using an anti-A22 Iraq, guinea-pig polyclonal sera. (B) The same ELISA using purified αvβ6-FLAG in place of transfected cell supernatants. (C) Wells were coated with cell culture supernatant from mock-transfected cells (Mock) or cells co-transfected with both αv and β6-FLAG subunits (αvβ6) and incubated with FMDV A22ecRGD (ecRGD) in the presence or absence of an RGD-containing peptide, an inactive control RGE peptide or EDTA (as indicated on the figure). (D) An ELISA using truncated αvβ6-FLAG to detect FMDV empty capsid. Wells were coated with A22ecRGD (RGD), A22ecKGA (KGA) or coating buffer alone (CB) and capsid proteins detected using FLAG-tagged truncated αvβ6 (αvβ6) i.e. cell culture supernatant from αvβ6 co-transfected cells. RGD/Mock wells were coated with A22ecRGD and incubated with cell culture supernatant from mock-transfected cells.