Fig 6. VgrG5-dependent fusion required for increased pathogenesis in GBP deficiency.

Unprimed BMDMs were infected (MOI 5) with B. thailandensis (WT or vgrG5ΔCTD) and (a) phase contrast images (20x) were taken after 20 h to observe the extent of MNGC fusion. (b) Immunoblots were probed for caspase-1 cleavage following infection with B. thailandensis (WT or vgrG5ΔCTD). (c) BMDMs were infected with B. thailandensis (WT or vgrG5ΔCTD) and intracellular bacterial CFUs were determined by serial dilution of lysed cells at the indicated time points. (d) Indicated mouse strains were inoculated (i.n.) with B. thailandensis (WT or vgrG5ΔCTD) (1 x 10⁵) and monitored for survival following infection for 7 days. (e,f) Wildtype and Gbp2^−/− mice were inoculated with B. thailandensis (WT or vgrG5ΔCTD, 5 x 10³) and lungs were fixed in formalin, stained with H&E, and scored (described in Methods) by a trained pathologist (PV). Statistical significance was determined by Dunnet’s multiple comparison test (c), the log-rank test (d), or one-way ANOVA with Tukey’s multiple comparison test (f), ****P < 0.0001. Data are representative of (a,b,c) at least three independent experiments, (d) pooled from two independent repeats, or (e,f) a single experiment.