Bindloss et al A B W日PagD 00000 0000 8s 0p0 奖mS要 oO § Strep: c17: Tributyrin: H ■ Isotype control ■ sotype control U Anti-CD25 口AncD25 ×3.5 9 control cD25 control CD25 8 WT Pparg WT Pparg u WT Pparg Figure 4. Microbiota-induced PPAR-y-signaling and Tregs cooperate to limit the bioavailability of oxygen in the colon (A and B)Groups of mice(N= 4)were treated with streptomycin(A)or with anti-CD2 antibody(B)and CD3t-enriched live colonic cells analyzed for expression of CD4 and FOXP3 by flow cytometry.( C) Groups of mice(N=6)were treated with anti-CD25 antibody or isotype control and 10 days later inoculated with a 1: I mixture of an avirulent Salmonella strain(invA spiB mutant) and an avirulent Salmonella strain lacking cytochrome bdl oxidase(inv A spiB cyxA mutant). (D)Groups(N= 6)of streptomycin-treated or mock treated mice were inoculated with Salmonella indicator strains and received supplementation with tributyrin or a community of 17 human Clostridia isolates( C17).(C and D). The CI was determined 4 days after inoculation(E-G)Groups of mice(N= 6) were treated with anti-CD25 antibody or isotype control antibody and colonocytes isolated to measure intracellular concentrations of lactate(E), ATP(F) or mitochondrial cytochrome c oxidase activity.(G) Boxes in Whisker plots represent the standard error, while lines indicate the standard deviation(A-B and E-F) Bars represent geometric means+ standard error (C, D and H)Each dot represents data from an individual animal and black bars represent geometric means. *, P<0.05; ** P<0.01,P<0.001; ns, not statistically significantly different Science Author manuscript; available in PMC 2017 October 1Figure 4. Microbiota-induced PPAR-γ-signaling and Tregs cooperate to limit the bioavailability of oxygen in the colon (A and B) Groups of mice (N = 4) were treated with streptomycin (A) or with anti-CD25 antibody (B) and CD3+-enriched live colonic cells analyzed for expression of CD4 and FOXP3 by flow cytometry. (C) Groups of mice (N = 6) were treated with anti-CD25 antibody or isotype control and 10 days later inoculated with a 1:1 mixture of an avirulent Salmonella strain (invA spiB mutant) and an avirulent Salmonella strain lacking cytochrome bdII oxidase (invA spiB cyxA mutant). (D) Groups (N = 6) of streptomycin-treated or mock￾treated mice were inoculated with Salmonella indicator strains and received supplementation with tributyrin or a community of 17 human Clostridia isolates (C17). (C and D). The CI was determined 4 days after inoculation. (E–G) Groups of mice (N = 6) were treated with anti-CD25 antibody or isotype control antibody and colonocytes isolated to measure intracellular concentrations of lactate (E), ATP (F) or mitochondrial cytochrome c oxidase activity. (G) Boxes in Whisker plots represent the standard error, while lines indicate the standard deviation. (A–B and E–F) Bars represent geometric means ± standard error. (C, D and H) Each dot represents data from an individual animal and black bars represent geometric means. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not statistically significantly different. Byndloss et al. Page 20 Science. Author manuscript; available in PMC 2017 October 16. Author Manuscript Author Manuscript Author Manuscript Author Manuscript