35 C Butyrate production 30 aPeptostreptococcaceae 815 g veillonellaceae 04 10 □ Ruminococcaceae IFNY/IL-22 口 Lachnospiraceae p (2 mM)5mM) E G H 816 i AiiA' 105 Strep: Strep Strep:-+ GW9662 +GW962 GW9662 GW9662 Figure 1. The PPAR-y agonist butyrate limits the availability of nitrate by repressing Nos2 expression (A)Streptomycin(Strep-treated mice(N=8)were inoculated with a 1: I mixture of E. coli wild type(wt) and napA narG narZ mutant and received rosiglitazone( rosi) or aminoguanidine(AG) supplementation. The competitive index (Cl; the ratio of wt and napA narG narZ mutant recovered from colon contents) was determined 3 days after inoculation B) Polarized Caco-2 cells(N=4)grown in tissue culture medium received IFNy/IL-22, Rosi or AG treatment. Nitrate produced in the apical compartment was determined by a modified Griess assay(C-F) Mice(N= 8)were mock-treated (inoculation with vehicle control)or treated with Strep and organs collected 3 days later.( C)The abundance of Clostridia in colon contents was determined by quantitative real-time PCR using class- specific primers for Clostridia 16S rRNA genes. (D)Relative abundance of families belonging to the class Clostridia determined by 16S profiling of DNA isolated from colon contents.(E)The butyrate concentration was determined in cecal contents using gas chromatography and (f)transcript level of Nos2 in colonocyte preparations was determined by real-time PCR. (G and H) Colonocytes were isolated 3 days after treatment with streptomycin from mice(N= 8)receiving the indicated supplementation and transcript levels of Angptl4(G)and Nos2 (H) determined by quantitative real-time PCR (I and D) Mice(N=6)were mock-treated or received the PPAR-y antagonist Gw9662 and were inoculated with E coli indicator strains. Numbers of E. coli and the cl of indicator strains()were determined 3 days after inoculation. (C and E-f) Bars represent geometric means+ standard error.(A, B and D) Dots represent measurements from individual animals (A and )or wells(B)and bars represent geometric means. P<0.05:,P<0.01 Science Author manuscript; available in PMC 2017 October 1Figure 1. The PPAR-γ agonist butyrate limits the availability of nitrate by repressing Nos2 expression (A) Streptomycin (Strep)-treated mice (N = 8) were inoculated with a 1:1 mixture of E. coli wild type (wt) and napA narG narZ mutant and received rosiglitazone (Rosi) or aminoguanidine (AG) supplementation. The competitive index (CI; the ratio of wt and napA narG narZ mutant recovered from colon contents) was determined 3 days after inoculation. (B) Polarized Caco-2 cells (N = 4) grown in tissue culture medium received IFNγ/IL-22, Rosi or AG treatment. Nitrate produced in the apical compartment was determined by a modified Griess assay. (C–F) Mice (N = 8) were mock-treated (inoculation with vehicle control) or treated with Strep and organs collected 3 days later. (C) The abundance of Clostridia in colon contents was determined by quantitative real-time PCR using class￾specific primers for Clostridia 16S rRNA genes. (D) Relative abundance of families belonging to the class Clostridia determined by 16S profiling of DNA isolated from colon contents. (E) The butyrate concentration was determined in cecal contents using gas chromatography and (F) transcript level of Nos2 in colonocyte preparations was determined by real-time PCR. (G and H) Colonocytes were isolated 3 days after treatment with streptomycin from mice (N = 8) receiving the indicated supplementation and transcript levels of Angptl4 (G) and Nos2 (H) determined by quantitative real-time PCR. (I and J) Mice (N = 6) were mock-treated or received the PPAR-γ antagonist GW9662 and were inoculated with E. coli indicator strains. Numbers of E. coli (I) and the CI of indicator strains (J) were determined 3 days after inoculation. (C and E–I) Bars represent geometric means ± standard error. (A, B and J) Dots represent measurements from individual animals (A and J) or wells (B) and bars represent geometric means. *, P < 0.05; **, P < 0.01. Byndloss et al. Page 17 Science. Author manuscript; available in PMC 2017 October 16. Author Manuscript Author Manuscript Author Manuscript Author Manuscript