Supplementary Figure 3: PAS638 senses tetrathionate in cybb^−/−, IL10^−/−, and 129X1/SvJ mouse models.

a) Weight of cybb-/- and C57Bl/6 control mice was measured following administration of PAS638 ± S. typhimurium (S.tm) ΔttrR. Graph shows percentage of pre-administration weight of individual mice (dotted) along with group averages (solid line). b) Elevated LCN-2 levels were apparent in S. typhimurium ΔttrR infected mice and cybb-/- uninfected controls. Graph shows maximum measurements from days 3-5 following bacterial administration. Means are marked. *p=0.01, **p=0.009, F(3,16) = 6.575, using one-way ANOVA analysis with Tukey’s multiple comparison correction. For all other comparisons p>0.99. c) LCN-2 levels were elevated in IL10-/- compared to control mice in the week following PAS638 administration therefore PAS638 measurements were taken >1-week following administration. Values are averages of 2-3 measurements from individual mice, with mean shown. *p=0.02, **p=0.01, F(2,20) = 6.48 using one-way ANOVA test with Tukey’s multiple comparison correction. >week 1 IL10-/- samples were not significantly increased over C57Bl/6 controls (p=0.9) d) A subset of IL10-/- mice administered PAS638 without streptomycin pre-treatment (n=10) showed elevated PAS638 switching. Graph shows maximum switching percentage from 4 measurements in the 12 days following administration. Mean is marked. e) When administered without streptomycin pre-treatment no indication of elevated LCN-2 levels was seen following bacterial administration. Graph shows pre-administration levels (1 measurement) and average levels from 4 measurements in the 12 days following administration. Means are marked. n.s. = non-significant (p = 0.6) using a two-tailed Mann-Whitney test. f). PAS638 administered to 129X1/SvJ mice (n=5) without streptomycin pre-treatment colonized the gut successfully. Graph shows CFU values from individual mice (dotted lines) and average across the group (solid line). g) The absence of streptomycin pre-treatment did not affect PAS638 switching in these mice. Graph shows average switching from 4 measurements in the 12 days following bacterial administration. h) Histology scoring of small intestine, cecum, proximal and distal colon showed no signs of overt inflammation in 18 129X1/SvJ mice, including all that showed considerable switching by PAS638.