Figure 1
Characterization of P. gingivalis ΔporA mutant. (A) Pigmentation of the ΔporA mutant on blood agar plates for 3 days. (B) Hemagglutinating activity of the ΔporA mutant. (C) Kgp and Rgp activities of the ΔporA mutant. The cell lysates (cell) and vesicle-containing culture supernatants (sup) were subjected to the assay. Lane 1: wild type, Lane 2: ΔporA, Lane 3: ΔporA/porA+. (D) Immunoblot analyses of T9SS CTD-containing proteins and A-LPS in the ΔporA mutant. Cell lysates of the wild type (lane 1), ΔporA (lane 2), and ΔporA/porA+ (lane3) were analyzed by SDS-PAGE, followed by immunoblot analyses of T9SS CTD-containing proteins and A-LPS using antibodies against PorA (α-PorA), Kgp (α-Kgp), Rgp (α-Rgp) and Hbp35 (α-Hbp35), and monoclonal antibody against A-LPS (mAb 1B5). (E) Cell lysates of the wild type (lane 1), ΔporA (lane 2), and ΔporA/porA+ (lane3) were analyzed by SDS-PAGE, followed by immunoblot analyses of T9SS component proteins using antibodies against PorU (α-PorU), PorK (α-PorK), PorL (α-PorL), PorM (α-PorM), and PorN (α-PorN). (F) Immunoblot analyses of the T9SS regulatory proteins SigP, PorY, and PorX in various T9SS-related mutants. Cells of P. gingivalis strains were lysed with 1% N-Dodecyl-β-d-maltopyranoside (DDM) (vol/vol) and then subjected to SDS-PAGE, followed by immunoblot analysis using antibodies against SigP (α-SigP), PorX (α-PorX), and PorY (α-PorY). Full-length blots/gels of (E) and (F) are presented in Supplementary Figure S11.
