Volume 22, Issue 6, Pages 971-982 (December 2015) Dietary Fiber-Induced Improvement in Glucose Metabolism Is Associated with Increased Abundance of Prevotella  Petia Kovatcheva-Datchary, Anne Nilsson, Rozita Akrami, Ying Shiuan Lee, Filipe De Vadder, Tulika Arora, Anna Hallen, Eric Martens, Inger Björck, Fredrik Bäckhed  Cell Metabolism  Volume 22, Issue 6, Pages 971-982 (December 2015) DOI: 10.1016/j.cmet.2015.10.001 Copyright © 2015 Elsevier Inc. Terms and Conditions

Cell Metabolism 2015 22, 971-982DOI: (10.1016/j.cmet.2015.10.001) Copyright © 2015 Elsevier Inc. Terms and Conditions

Figure 1 Glucose and Insulin Profiles in Non-responders and Responders after a Standardized Breakfast following 3-Day Consumption of WWB or BKB (A–E) Blood glucose and serum insulin responses (A–D) and fasting breath hydrogen (H2) (E) in non-responders (n = 10) and responders (n = 10) following a standardized breakfast after 3-day consumption of WWB or BKB. Data are mean ± SEM (∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; and ∗∗∗∗p < 0.0001; Student’s t test). Related to Figures S1 and S3 and Table S1. Cell Metabolism 2015 22, 971-982DOI: (10.1016/j.cmet.2015.10.001) Copyright © 2015 Elsevier Inc. Terms and Conditions

Figure 2 Altered Gut Microbiota in Non-responders and Responders following 3-Day Consumption of WWB or BKB (A) Weighted UniFrac distances in non-responders (n = 10) and responders (n = 10) at baseline and after 3-day consumption of WWB and BKB based on bacterial V1-V2 16S rRNA pyrosequencing data. Data are mean ± SEM (∗p < 0.05; ∗∗∗p < 0.001; Student’s t test with 1,000 Monte Carlo simulations). (B) Relative abundance of the major microbial phyla in non-responders and responders at baseline and after 3-day consumption of WWB and BKB (∗p < 0.05 baseline versus BKB; Wilcoxon matched-pairs signed rank test corrected for FDR). (C and D) Bacteroidetes phyla (C) and Prevotella/Bacteroides (D) ratio in non-responders and responders at baseline and after 3-day consumption of WWB and BKB. Data are mean ± SEM (∗p < 0.05; Wilcoxon matched-pairs signed rank test; ap = 0.056; Mann-Whitney U test). Related to Figure S2. Cell Metabolism 2015 22, 971-982DOI: (10.1016/j.cmet.2015.10.001) Copyright © 2015 Elsevier Inc. Terms and Conditions

Figure 3 Taxonomic Composition of the Metagenome of Non-responders and Responders at Baseline and after BKB Consumption (A) Microbiota composition (family level) of non-responders and responders at baseline and after BKB. The ten top abundant families are shown. Data are mean ± SD. (B) Prevotella/Bacteroides ratio. Data are mean ± SD (∗p < 0.05; Wilcoxon matched-pairs signed rank test; ap = 0.056; Mann-Whitney U test). (C) Prevotella composition (species level) with total read abundance above 0.01. Data are mean ± SD. For statistics data, see Table S2. Related to Figure S4. Cell Metabolism 2015 22, 971-982DOI: (10.1016/j.cmet.2015.10.001) Copyright © 2015 Elsevier Inc. Terms and Conditions

Figure 4 Beneficial Effects of Prevotella on Glucose Metabolism in Mice Are Diet Dependent, and P. copri Protects against B. thetaiotaomicron-Induced Glucose Intolerance (A and B) Oral glucose tolerance test in (A) Swiss Webster and (B) C57BL/6 mice fed standard chow diet and gavaged with heat-killed P. copri or live P. copri for 7 days. Data are mean ± SEM (∗p < 0.05, ∗∗p < 0.01; Student’s t test). (C) Oral glucose tolerance test in C57BL/6 mice fed high-fat diet and gavaged with P. copri or media for 7 days. Data are mean ± SEM. (D) Levels of B. thetaiotaomicron and P. copri determined by 16S rRNA specific qPCR following 14-day colonization of GF mice. Data are mean ± SEM (∗p < 0.05; Mann-Whitney U test). (E) Oral glucose tolerance test following 14-day colonization of GF mice with B. thetaiotaomicron, P. copri, or both strains in Swiss Webster mice fed chow diet. Data are mean ± SEM (∗p < 0.05; ∗∗p < 0.01; Student’s t test; ap < 0.05 BT versus BT+P; Student’s t test). Related to Figure S5. Cell Metabolism 2015 22, 971-982DOI: (10.1016/j.cmet.2015.10.001) Copyright © 2015 Elsevier Inc. Terms and Conditions

Figure 5 Transfer of a Responder’s Gut Microbiota Taken after BKB to GF Mice Improves Glucose Tolerance (A and B) Oral glucose tolerance test in recipient Swiss Webster male mice colonized with (A) non-responder donor 1 or (B) responder donor 1 microbiota taken at baseline and after BKB. Data are mean ± SEM (∗p < 0.05; Student’s t test). (C and D) Levels of (C) Prevotella and (D) Bacteroides in the cecum of recipient mice after 14 days of colonization, determined by 16S rRNA specific qPCR. Data are mean ± SEM (∗p < 0.05; ∗∗p < 0.01; Mann-Whitney U test). (E) Prevotella to Bacteroides ratio. Data are mean ± SEM (∗∗p < 0.01; ∗∗∗∗p < 0.0001; Mann-Whitney U test). Related to Figures S6 and S7. Cell Metabolism 2015 22, 971-982DOI: (10.1016/j.cmet.2015.10.001) Copyright © 2015 Elsevier Inc. Terms and Conditions

Figure 6 Transfer of a Responder’s Gut Microbiota Taken after BKB to GF Mice Promotes Hepatic Glucose Storage as Glycogen (A–C) Relative expression of (A) G6pc in proximal colon, (B) Sorbs1 in liver, and (C) Pyg1 in liver of recipient mice colonized with non-responder or responder microbiota taken at baseline and after BKB. Data are mean ± SEM (∗p < 0.05;∗∗p < 0.01; Mann-Whitney U test). (D) Glycogen in the liver of recipient mice colonized with non-responder or responder microbiota taken at baseline and after BKB. Data are mean ± SEM (∗∗∗∗p < 0.0001; ANOVA with Tukey post-hoc test). Cell Metabolism 2015 22, 971-982DOI: (10.1016/j.cmet.2015.10.001) Copyright © 2015 Elsevier Inc. Terms and Conditions