Journal: PLoS ONE

Article Title: Trimeric Form of Intracellular ATP Synthase Subunit β of Aggregatibacter actinomycetemcomitans Binds Human Interleukin-1β

doi: 10.1371/journal.pone.0018929

Figure Lengend Snippet: The tested strains D7S (serotype a) was rough colony forming isolate, which produced fimbriae. Pre-grown (approximately 18 h) biofilms were grown with human IL-1β (10 ng/ml) or without (control) in RPMI 1640 medium and the metabolic activity was followed with alamarBlue™ (Panel A). Data is shown as percentage of reduced alamarBlue compared to control. The spontaneous smooth-colony variant (D7SS) of D7S and its planktonic single-gene deletion mutants rcpA , rcpB , tadD , tadG , and flp1-flp2 were studied (Panel B and C). The pre-grown (18 h) cells were incubated with or without human IL-1β (10 ng/ml) in RPMI 1640 medium, and the metabolic activity was followed using alamarBlue™. The results are shown as percentage of metabolic activities of IL-1β-containing reactions compared to control cultures (Panel B). The metabolic activities of planktonic control cultures (sterile water was substituted for IL-1β) are presented as the fluorescence of the reduced form of alamarBlue™ (Panel C). All results are shown as means ± SD from three independent experiments. Each experiment contained duplications of each reaction.

Article Snippet: The endogenous peroxidase block and protein block treatments were done before overnight incubation with rabbit anti-IL-1β antibody (Novus Biologicals, Littleton, CO, USA), control rabbit IgG whole molecule (Jackson ImmunoResearch Laboratories, PA, USA), or control rabbit anti-N-terminal-RcpA antibody (ABCELL, Tampere, Finland) at 4°C.

Techniques: Produced, Control, Activity Assay, Variant Assay, Incubation, Sterility, Fluorescence

Journal: PLoS ONE

Article Title: Trimeric Form of Intracellular ATP Synthase Subunit β of Aggregatibacter actinomycetemcomitans Binds Human Interleukin-1β

doi: 10.1371/journal.pone.0018929

Figure Lengend Snippet: Formalin fixed paraffin sections of A. actinomycetemcomitans biofilm containing co-cultures were treated with anti-IL-1β (Panel A), anti-N-terminal-RcpA (Panel B), or control IgG (Panel C) after which the binding antibodies were detected with the NovoLink™ Polymer Detection System (Novocastra™). The sections with the DAB-label were stained with osmium for electron microscopy. Anti-IL-1β stained samples showed structures of A. actinomycetemcomitans cell shape and size (Panel D), with dark precipitate in both extra- and intracellular space (Panel G). The anti-RcpA stained positive control showed intense staining (Panel B) with similar structures (Panel E), although the cell structures are less visible due to the extent of the extracellular precipitate (Panel H). Control IgG antibody showed less staining (Panel C) revealing similar structures (Panel F) without dark precipitates bound to the cell membranes (Panel I).

Article Snippet: The endogenous peroxidase block and protein block treatments were done before overnight incubation with rabbit anti-IL-1β antibody (Novus Biologicals, Littleton, CO, USA), control rabbit IgG whole molecule (Jackson ImmunoResearch Laboratories, PA, USA), or control rabbit anti-N-terminal-RcpA antibody (ABCELL, Tampere, Finland) at 4°C.

Techniques: Control, Binding Assay, Polymer, Staining, Electron Microscopy, Positive Control

Journal: PLoS ONE

Article Title: Trimeric Form of Intracellular ATP Synthase Subunit β of Aggregatibacter actinomycetemcomitans Binds Human Interleukin-1β

doi: 10.1371/journal.pone.0018929

Figure Lengend Snippet: Aggregatibacter actinomycetemcomitans strains used in this study.

Article Snippet: The endogenous peroxidase block and protein block treatments were done before overnight incubation with rabbit anti-IL-1β antibody (Novus Biologicals, Littleton, CO, USA), control rabbit IgG whole molecule (Jackson ImmunoResearch Laboratories, PA, USA), or control rabbit anti-N-terminal-RcpA antibody (ABCELL, Tampere, Finland) at 4°C.

Techniques: