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. 2017 Jul 24;13(7):e1006531. doi: 10.1371/journal.ppat.1006531

Fig 5. Vag8 interferes with the binding of C1s, C1r and MASP-2 with C1-inh.

Fig 5

(A) B. pertussis strain B1917 was opsonized with 1.25% NHS with or without Vag8 or Prn. Supernatant was analysed using anti C1-inh. The presence of Vag8 results in a decreased signal of the ~150 kDa band. (B) In a purified system consisting of C1q, C1s or C1r with C1-inh in the presence or absence of Vag8 or Prn, we show, using anti-C1-inh, that C1s and C1r, but not C1q, bind to C1-inh and that this binding is inhibited by the addition of Vag8 but not Prn. (C) C1r was detected using anti-C1r. The presence of C1r bound to C1 and C1-inh is inhibited in the presence of Vag8 but not Prn. Additionally, we show the presence of active C1r in the presence of Vag8. (D) Purified MASP-2 was incubated with C1-inh in the presence of Vag8 or Prn and analyzed using anti-C1-inh. We show inhibition of MASP-2 binding to C1-inh in the presence of Vag8, but not Prn. Immunoblots are representative of three separate experiments.