The replication kinetics of the galU mutant within J774 or RAW 264.7 cells were indistinguishable from those of the WT strain (Figure 1C), indicating that mutation of the galU gene had no effect on uptake or intracellular survival/replication of the bacterium. Virulence of the galU mutant in vivo To determine whether the galU gene is important for FT virulence, C57Bl/6J mice (5/group) were inoculated intranasally with 5 × 104 CFU (50 × LD50) of either selleck compound the galU mutant or WT FT and then were monitored for 15 days. Each of the mice challenged with the galU mutant experienced transient weight loss but

survived and completely cleared the infection, while all of the mice challenged with WT FT lost weight continually until they succumbed to tularemia (Figure 2A and

2B). An additional {Selleck Anti-diabetic Compound Library|Selleck Antidiabetic Compound Library|Selleck Anti-diabetic Compound Library|Selleck Antidiabetic Compound Library|Selleckchem Anti-diabetic Compound Library|Selleckchem Antidiabetic Compound Library|Selleckchem Anti-diabetic Compound Library|Selleckchem Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|buy Anti-diabetic Compound Library|Anti-diabetic Compound Library ic50|Anti-diabetic Compound Library price|Anti-diabetic Compound Library cost|Anti-diabetic Compound Library solubility dmso|Anti-diabetic Compound Library purchase|Anti-diabetic Compound Library manufacturer|Anti-diabetic Compound Library research buy|Anti-diabetic Compound Library order|Anti-diabetic Compound Library mouse|Anti-diabetic Compound Library chemical structure|Anti-diabetic Compound Library mw|Anti-diabetic Compound Library molecular weight|Anti-diabetic Compound Library datasheet|Anti-diabetic Compound Library supplier|Anti-diabetic Compound Library in vitro|Anti-diabetic Compound Library cell line|Anti-diabetic Compound Library concentration|Anti-diabetic Compound Library nmr|Anti-diabetic Compound Library in vivo|Anti-diabetic Compound Library clinical trial|Anti-diabetic Compound Library cell assay|Anti-diabetic Compound Library screening|Anti-diabetic Compound Library high throughput|buy Antidiabetic Compound Library|Antidiabetic Compound Library ic50|Antidiabetic Compound Library price|Antidiabetic Compound Library cost|Antidiabetic Compound Library solubility dmso|Antidiabetic Compound Library purchase|Antidiabetic Compound Library manufacturer|Antidiabetic Compound Library research buy|Antidiabetic Compound Library order|Antidiabetic Compound Library chemical structure|Antidiabetic Compound Library datasheet|Antidiabetic Compound Library supplier|Antidiabetic Compound Library in vitro|Antidiabetic Compound Library cell line|Antidiabetic Compound Library concentration|Antidiabetic Compound Library clinical trial|Antidiabetic Compound Library cell assay|Antidiabetic Compound Library screening|Antidiabetic Compound Library high throughput|Anti-diabetic Compound high throughput screening| challenge trial in which C57Bl/6 mice (4/group) were challenged with higher numbers of the galU mutant (up to 107 CFU) revealed that this mutant is highly attenuated, with an LD50 that is at least 5 logs higher than that of WT FT (Figure 2C). Moreover, trans-complementation of the galU mutation completely restored virulence of the mutant strain (Figure 2A). These findings indicated that FT virulence in mice is dependent on the expression of a functional galU gene product. Figure 2 Mutation of the galU gene Histone Methyltransferase inhibitor attenuates virulence of FT. C57BL/6 mice were infected intranasally with 5 × 104 CFU of WT (n = 9), the galU mutant (n = 10), or the galU-complemented strain (n = 5) strain of FTLVS, and their survival (Panel A) and weight (Panel B) were monitored. Statistical analyses of survival curves was performed using Gehan-Breslow-Wilcoxon tests and a p value of 0.005

is indicated (**). Statistical analysis of body weight retention was performed via one-way ANOVA with a Bonferroni multiple comparisons post-test and a p value of <0.0001 is indicated (***). Panel C: Survival was also monitored in C57Bl/6J mice challenged with a range of higher doses of the galU mutant (1 × 105-1 × 107 CFU; n = 4) or WT FT (5 × 104 CFU; n = 5). Statistical analysis of survival curves was performed using Gehan-Breslow-Wilcoxon many tests and p values of 0.027 (*) and 0.009 (**) are indicated. Results shown are representative of two experiments of similar design. To determine whether the reduced virulence of the galU mutant was the result of defective replication and/or dissemination of the bacterium in vivo, we performed a kinetic analysis of bacterial burdens following infection. C57Bl/6J mice (16/group) were challenged with 5 × 104 CFU of either the galU mutant or WT FT and then four mice were sacrificed at each time point (24, 48, 72, and 96 h post-infection) for bacterial burden determinations from the lungs, livers, and spleens (Figure 3).