Figure 9. Genetic mutation of HAd-Ft confers a pro-inflammatory phenotype upon the bacterium.

Primary BMDMs (2.5×10⁵ cells/well) were infected with wild-type and mutant HAd-Ft at a MOI of 100. Supernatants were collected 24 h p.i. and were assayed for the presence of (A) TNF, (B) IL-1β, and (C) IL6 by CBA. Results represent the mean ± SEM from four independent experiments. **P<0.01, ***P<0.001. (D) HAd-Ft and the corresponding mutants were stained with SYTOX® Green nucleic acid stain and were analyzed for SYTOX⁺ bacteria by flow cytometry. Results represent the mean ± SEM from three independent experiments. **P<0.01, ***P<0.001. (E) Primary BMDMs (2.5×10⁵ cells/well) were infected with wild-type and mutant HAd-Ft at a MOI of 100. After gentamycin treatment for 2 h, intra and extracellular bacteria were recovered at 4 and 24 h and enumerated by colony plating. Results represent the mean ± SEM from three independent experiments. *P<0.05, **P<0.01. (All results shown were subjected to One-way ANOVA with Bonferroni's Post-test).