Figure 4 | Scientific Reports

Figure 4

From: Immune evasion of Borrelia miyamotoi: CbiA, a novel outer surface protein exhibiting complement binding and inactivating properties

Figure 4

Borrelial proteins inhibit TCC deposition of the CP and TP. In order to analyze the inhibitory effect of borrelial proteins on the AP (A), CP (B), and TP (C) a hemolytic assay was employed. Amboceptor-sensitized sheep erythrocytes (CP) and rabbit erythrocytes (AP) were incubated with NHS or with NHS pre-incubated with purified proteins or BSA in either Mg-EGTA buffer (AP) or GVB++ buffer (CP). Inhibition of the TP was investigated using sheep erythrocytes pre-incubated with the C5b-6 complex. A reaction mixture containing C7, C8, and C9 was pre-incubated with increasing concentrations of purified proteins or BSA. Following incubation, erythrocyte lysis was detected at 414 nm. Means from three separate experiments are shown and error bars correspond to SD. Raw data were analyzed using one-way ANOVA (A and B). ***P < 0.001, **P < 0.01, *P < 0.05. Binding of C9 to CbiA (D). C9 was labeled with NT-647 RED-NHS (NanoTemper technologies) and the interaction with CbiA was assessed in the fluid phase. The relative fluorescence in the thermophoresis phase has been plotted against the concentration of CbiA. The data shown are representative of three independent experiments.

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