FIG. 10.

Ectopic expression of BpcA by B. burgdorferi B313 isolate. (A) Expression of BpcA by transformed B. burgdorferi B313 was assessed using immunoblot analysis. Whole-cell lysates were separated by 10% SDS-PAGE, transferred to nitrocellulose, and analyzed for CFH binding by incubation with NHS and a CFH-specific MAb (JHD-7). (B) Surface localization of BpcA by transformed B. burgdorferi B313 cells was assessed using trypsin and proteinase K treatment. B313/pBP cells were incubated with the indicated concentrations of trypsin, lysed, immunoblotted, and probed with either a BpcA-specific MAb, BP3, or antiflagellin MAb LA21 (lower panel). (C) To investigate susceptibility to human serum, transformed B313/pBP or B313 cells were incubated with 25% NHS at 30°C for 72 h. Cells were stained with a nucleic acid dye, and the relative growth compared to day 0 levels was determined by measurement of green fluorescence emission. Values represent the means ± SEM of results from a single experiment performed in triplicate that is representative of three independent experiments. ***, P < 0.0002.