a118  (ATCC)

Journal: eLife

Article Title: Capsular polysaccharide restrains type VI secretion in Acinetobacter baumannii

doi: 10.7554/eLife.101032

Figure Lengend Snippet: ( A ) Illustration of the T6SS gene arrangement in A. baumannii strain A118. The core components of the T6SS are highlighted in gray. ( B ) Survival of E. coli following interaction with wild-type (WT) (strain A118), the two T6SS-inactive mutants (Δ hcp and Δ tssB ), or the TssB translational fusion-carrying strain ( tssB-msfGFP ) of A. baumannii . Survival rates are presented on the Y -axis. ( C, D ) A. baumannii kills E. cloacae in a T6SS-dependent manner. ( C ) Survival of capsulated (WT), T6SS-inactive (Δ hcp ), non-capsulated (Δ itrA ), or T6SS-inactive and non-capsulated (Δ hcp Δ itrA ) A. baumannii after co-incubation with T6SS+ (white bars) or T6SS− (dashed bars) E. cloacae . Survival rates for E. cloacae are shown as in panel ( D ). Data points are from three independent experiments, with bars indicating mean values (± SD, depicted by error bars). Statistical significance was determined using an ordinary one-way ANOVA test. *p < 0.05, **p < 0.01, ****p < 0.0001, ns = not significant. The detection limit (dl) is indicated.

Article Snippet: Antibody , Anti Hcp-A118-2 polyclonal antibodies (produced in rabbit) , Eurogentec , Custom-made ( 2110850 cl.2 ) , 1:667 diluted.

Techniques: Incubation

Journal: eLife

Article Title: Capsular polysaccharide restrains type VI secretion in Acinetobacter baumannii

doi: 10.7554/eLife.101032

Figure Lengend Snippet: ( A , B ) Colony morphologies on blood agar plates after 24 hr of growth, with strain genotypes indicated. ( B ) depicts zoomed regions of the white boxes shown in panel A. ( C ) Complementation of bfmS deletion assessed by T6SS-mediated killing activity. Enumeration of E. coli after exposure to wild-type (WT), T6SS-inactive (Δ hcp ) , Δ bfmS , and the bfmS -complemented strain (Δ bfmS::bfmS ), with survival shown on the Y -axis. ( D ) Capsule production in WT, capsule-deficient (Δ itrA ), capsule-overproducing (Δ bfmS ), and secretion-impaired (Δ tssB ) strains. Polysaccharides from supernatant were precipitated, separated by SDS–PAGE, and visualized with Alcian blue staining. The arrowhead marks the polysaccharide band. ( E ) Hcp secretion remains detectable in WT co-cultured with secretion-impaired (+Δ tssB ) or capsule-overproducing ∆ bfmS (+Δ bfmS ) strains, as analyzed by immunoblotting. Details as described in . ( F ) Survival of E. coli prey after contact with A. baumannii WT and mutants Δ hcp, Δ bfmS strains across various strain backgrounds (A118, 29D2, and 86II/2C). Details as in panel C. Statistical analysis as in with ****p < 0.0001. Figure 3—figure supplement 1—source data 1. PDF file containing the original gels for , indicating the relevant bands. Figure 3—figure supplement 1—source data 2. Original files for gel displayed in . Figure 3—figure supplement 1—source data 3. PDF file containing original western blots for , indicating the relevant bands and treatments. Figure 3—figure supplement 1—source data 4. Original files for western blot analysis displayed in .

Article Snippet: Antibody , Anti Hcp-A118-2 polyclonal antibodies (produced in rabbit) , Eurogentec , Custom-made ( 2110850 cl.2 ) , 1:667 diluted.

Techniques: Activity Assay, SDS Page, Staining, Cell Culture, Western Blot

Journal: eLife

Article Title: Capsular polysaccharide restrains type VI secretion in Acinetobacter baumannii

doi: 10.7554/eLife.101032

Figure Lengend Snippet: ( A ) Hcp levels in wild-type (WT) and the Δ tssB mutant of A. baumannii strains A118, 29D2, and 86IIC were evaluated under exponential (left) and stationary (right) growth phases via immunoblot analysis. Details as described for . ( B ) TssB production remains equal during stationary phase. Strains harboring the translational fusion TssB-msfGFP were cultured under stationary growth conditions and analyzed for GFP production. Details on strains and immunoblotting conditions as described for panel . ( C ) Growth curve of WT and the Δ tssB mutant over a 16-hr timeframe, with the gray zone highlighting the observed period of Hcp degradation shown in . Figure 6—figure supplement 1—source data 1. PDF file containing original western blots for , indicating the relevant bands and treatments. Figure 6—figure supplement 1—source data 2. Original files for western blot analysis displayed in . Figure 6—figure supplement 1—source data 3. PDF file containing original western blots for , indicating the relevant bands and treatments. Figure 6—figure supplement 1—source data 4. Original files for western blot analysis displayed in .

Article Snippet: Antibody , Anti Hcp-A118-2 polyclonal antibodies (produced in rabbit) , Eurogentec , Custom-made ( 2110850 cl.2 ) , 1:667 diluted.

Techniques: Mutagenesis, Western Blot, Cell Culture

Journal: eLife

Article Title: Capsular polysaccharide restrains type VI secretion in Acinetobacter baumannii

doi: 10.7554/eLife.101032

Figure Lengend Snippet:

Article Snippet: Antibody , Anti Hcp-A118-2 polyclonal antibodies (produced in rabbit) , Eurogentec , Custom-made ( 2110850 cl.2 ) , 1:667 diluted.

Techniques: Clone Assay, Recombinant, Sequencing, Produced