BB0347 is surface exposed in <i>B. burgdorferi</i> MI-16.

<p><b>A)</b> Intact spirochetes were treated with different proteases for 30 min or 2 hrs. Western blots were run against whole cell lysates after these treatments and blotted with αBB0347, αOspC, or αFlaB. After two hours, no difference was observed between the control and protease-treated spirochetes in the αFlaB-blotted membranes, but OspC and BB0347 were almost completely degraded (Key: C: No protease control, K: proteinase K, P: pronase, T: trypsin). <b>B)</b> Intact bacteria were coated onto glass slides and fixed with 10% Formalin. Antibodies against the same proteins listed in (A) were used to stain and a secondary antibody conjugated with Dylight 488 was used for detection. DAPI was used as a secondary stain to localize spirochetes (see Materials and Methods). An additional control in which the <i>B. burgdorferi</i> membrane was compromised by desiccation, was included to verify that αFlaB antibodies were effective on fixed spirochetes. Key: Panel 1) αOspC, 2) αBB0347, 3) αFlaB 4) αFlaB with membrane disruption. BB0347 was detected in intact spirochetes, further verifying the surface exposure. White bars indicate a length of 10 µM, and the magnification is 1000×. Results are indicative of three independent experiments with similar outcomes.