Electronic supplementary CH5183284 datasheet material Additional file 1: Comparison of HmuY homologues. Ivacaftor Comparison of homologous HmuY amino-acid sequences identified in human pathogens (A) and bacteria identified in oral tissues (B). Amino-acid sequences lacking signal peptides are shown. Positions with identical amino acids in more than 30% of the sequences are shown in black boxes and partial homology is indicated in grey boxes. Phylogenetic relationship between homologous HmuY amino-acid sequences (C). Bacteria infecting the oral cavity are shown in bold. The phylogenetic tree was determined with the Neighbor-Joining method. Bootstrap values are included. Pgi, Porphyromonas gingivalis; Pen, P. endodontalis;

Pue, P. uenonis; Bfr, Bacteroides fragilis; Bfi, B. finegoldii; Bco, B. coprocola;

Bst, B. stercoris; Bdo, B. dorei; Bvu, B. vulgatus; Bov, B. ovatus; Bca, B. caccae; Bth, B. thetaiotaomicron; Bcp, B. coprophilus; Bsp, Bacteroides sp.; Coc, Capnocytophaga ochracea; Cgi, C. gingivalis; Csp, C. sputigena; Lbo, Leptospira borgpetersenii; Lin, L. interrogans; Ssp, Sphingobacterium spiritivorum; Pbi, Prevotella bivia; Por, P. oris; Pbe, P. bergensis; Pti, P. timonensis; Pme, P. melaninogenica; Pve, P. veroralis; Psp, Prevotella sp.; Pta, P. tannerae. www.selleckchem.com/products/LY2603618-IC-83.html (DOC 318 KB) Additional file 2: Analysis of surface exposure of HmuY. Analysis of surface exposure of P. gingivalis HmuY analyzed by whole-cell ELISA. P. gingivalis wild-type (A7436, W83) and hmuY deletion mutant (TO4) strains were grown in basal medium supplemented with hemin (Hm) or dipyridyl (DIP). The cells were washed and diluted with PBS (starting at OD660 = 1.0). Varying dilutions of P. gingivalis cells were adsorbed on the wells of the microtiter plate and reacted with pre-immune serum (A) or purified pre-immune IgGs (pre) (B) and immune anti-HmuY much serum (A) or purified immune anti-HmuY IgGs (im) (B). Representative data are shown. (DOC 74 KB) Additional file 3: P. gingivalis growth in broth cultures and biofilms, and biofilm accumulation. P. gingivalis growth was analyzed by measuring the OD at 660 nm, cell viability by plating cells on ABA

plates and colony forming unit (CFU) calculation, and biofilm accumulation by microtiter plate assay. (DOC 36 KB) References 1. Pihlstrom BL, Michalowicz BS, Johnson NW: Periodontal diseases. Lancet 2005, 366:1809–1820.PubMedCrossRef 2. Schenkein HA: Host responses in maintaining periodontal health and determining periodontal disease. Periodontol 2000, 200640:77–93. 3. Mayrand D, Holt SC: Biology of asaccharolytic black-pigmented Bacteroides species. Microbiol Rev 1988, 52:134–152.PubMed 4. Lamont RJ, Chan A, Belton CM, Izutsu KT, Vasel D, Weinberg A: Porphyromonas gingivalis invasion of gingival epithelial cells. Infect Immun 1995, 63:3878–3885.PubMed 5. Belton CM, Izutsu KT, Goodwin PC, Park Y, Lamont RJ: Fluorescence image analysis of the association between Porphyromonas gingivalis and gingival epithelial cells.