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. 2017 Sep 19;8:1145. doi: 10.3389/fimmu.2017.01145

Figure 3.

Figure 3

Assembly of the alternative pathway C3 convertase on factor H-related (FHR)-B. (A) Assembly of the C3bBb convertase on FHR-B. 5 µg/mL recombinant FHR-B, human FHR-4B (FHR-4), C3b as positive control and BSA as negative control were immobilized in microplate wells. After blocking, the wells were incubated with 10 µg/mL C3b for 1 h at 37°C. The alternative pathway C3 convertase was built up by adding purified factor B (FB), factor D (FD), and factor P (FP) for 30 min. The convertase was detected using a polyclonal anti-FB antiserum. Data are mean absorbance values + SD derived from four independent experiments. ***p < 0.001, one-way ANOVA. (B) Activity of the FHR-B bound convertase was measured by adding 10 µg/mL C3 to the wells for 1 h at 37°C. C3a generation was measured by Quidel’s C3a ELISA kit. Data are mean absorbance values + SD derived from four independent experiments. ***p < 0.001, one-way ANOVA.