Skip to main content
. 2019 Nov 26;10:2722. doi: 10.3389/fimmu.2019.02722

Figure 3.

Figure 3

Identification and surface exposure of FH/FHL-1-binding proteins in B. mayonii MN14-1420. (A,B) Detection of FH/FHL-1-binding proteins by Far Western blot analysis using NHS as source of FH and purified FHL-1 (750 ng/ml). Cell lysates obtained from B. mayonii MN14-1420, B. burgdorferi LW2, B. burgdorferi B31, B. burgdorferi PKa-1, B. garinii G1, and transformant G1/pCspA_Bmayo were separated by 10% Tris/tricine-SDS-PAGE and transferred onto a nitrocellulose membrane. Flagellin (FlaB) was detected with the monoclonal antibody L41 1C11. The FH-binding proteins (A) were visualized by applying an anti-FH antiserum and FHL-1-binding proteins (B) were detected by using an anti-CCP1-4 antiserum. The corresponding to CspA protein of B. mayonii (Bm) MN14-1420, CspA of B. burgdorferi (Bb) LW2, CspZ, ErpP, and ErpA of B. burgdorferi s.s. are indicated at the right. (C) in situ protease accessibility assay. Native spirochetes were incubated with or without proteinase K or trypsin, then lysed by sonication and total proteins were separated by 10% Tris/tricine-SDS-PAGE. The band corresponding to CspA of B. mayonii is indicated on the right. The mobilities of molecular mass standards are indicated on the left. A full scan of the original membranes is presented in Supplementary Figure 5.