The NLR Protein NLRP6 Does Not Impact Gut Microbiota Composition Paul Lemire, Susan J. Robertson, Heather Maughan, Ivan Tattoli, Catherine J. Streutker, Jaye M. Platnich, Daniel A. Muruve, Dana J. Philpott, Stephen E. Girardin  Cell Reports  Volume 21, Issue 13, Pages 3653-3661 (December 2017) DOI: 10.1016/j.celrep.2017.12.026 Copyright © 2017 The Authors Terms and Conditions

Cell Reports 2017 21, 3653-3661DOI: (10.1016/j.celrep.2017.12.026) Copyright © 2017 The Authors Terms and Conditions

Figure 1 NLRP6 Genotype Does Not Influence Microbial Community Structure in Littermates (A–C) Bray-Curtis PCoA plots for fecal (A), colon (B), and ileum (C) samples. (D–F) Relative abundances of bacterial taxa in WT and Nlrp6−/− littermates in feces (D), colon (E), and ileum (F). (G–O) Alpha diversity measures: Chao 1 (G–I), Shannon (J–L), and inverse Simpson (M–O) in the three gut compartments of WT and Nlrp6−/− mice. Each group represents an average value of 10 mice. Data are represented as mean ± SEM; p < 0.05 (Adonis) indicates statistically significant differences between WT and Nlrp6−/− mice. See also Figure S1. Cell Reports 2017 21, 3653-3661DOI: (10.1016/j.celrep.2017.12.026) Copyright © 2017 The Authors Terms and Conditions

Figure 2 Relative Abundance of Specific Bacterial Taxa in Littermates Is Not Altered by NRLP6 Genotype (A–O) Relative abundances of specific bacterial taxa in feces (A, D, G, J, and M), colon (B, E, H, K, and N), and ileum (C, F, I, L, and O) gut compartments of WT and Nlrp6−/− mice. Each group represents an average value of 10 mice. Data are represented as median ± interquartile range; p < 0.05 (Mann-Whitney or Kruskal-Wallis test) indicates statistically significant differences between WT and Nlrp6−/− mice. Cell Reports 2017 21, 3653-3661DOI: (10.1016/j.celrep.2017.12.026) Copyright © 2017 The Authors Terms and Conditions

Figure 3 Genotype-Independent Sex Effect Revealed in Single-Caged Littermates (A–L) Bray-Curtis PCoA plot, Chao 1 measure of alpha diversity, relative abundances of bacterial taxa, and LEfSe plots of discriminating taxa in feces (A–D), colon (E–H), and ileum (I–L) gut compartments. Each group represents an average value of 5–8 mice. Alpha diversity data are represented as mean ± SEM. PCoA p < 0.05 (Adonis) indicates statistically significant differences between WT and Nlrp6−/− mice or between male and female mice. See also Figures S1 and S2. Cell Reports 2017 21, 3653-3661DOI: (10.1016/j.celrep.2017.12.026) Copyright © 2017 The Authors Terms and Conditions

Figure 4 NLRP6 Genotype Does Not Influence DSS-Induced Colitis Nlrp6−/− and WT mice were unchallenged (water) or challenged with 2.5% DSS for 5 days in drinking water followed by 15 days of recovery with normal water. (A–C) Weight curves (A), DAI score (B), and measurement of colon length (C). (D) Histopathological evaluation of inflammation-associated parameters of animals unchallenged or challenged with DSS at day 7 (5 days DSS + 2 days normal water). The numbers on the left side of the image are as follows: 1, total area affected; 2, area severely destructed; 3, area severely affected; 4, destruction of mucosal architecture; 5, degree of cellular infiltration; 6, muscle thickening; 7, crypt abscess; and 8, goblet cell depletion. (E and F) Measurement of colon length (E) and representative H&E micrographs of colons of animals unchallenged or challenged with DSS at day 7 (F). (G–K) qPCR analysis of Il6 (G), Tnf, (H), Cxc1 (I), Il10 (J), and C3 (K) in colonic tissues of animals unchallenged or challenged with DSS at day 7. (L and M) IL-18 (L) and IL-1β (M) production from colon explant of animals unchallenged or challenged with DSS at day 7. (N) Levels of fecal lipocalin-2 at days 0, 3, 5, and 7 during DSS treatment. (O and P) Myeloperoxidase (MPO) quantification (O) and neutrophil count (P) in colonic tissues of animals unchallenged or challenged with DSS at day 7. Each group represents an average value of 18–20 mice obtained in three different experiments. Data are represented as mean ± SEM; ∗p < 0.05 and ∗∗p < 0.01 (one-way ANOVA) indicate statistically significant differences between unchallenged or DSS-treated mice or between WT and Nlrp6−/− mice. Scale bar, 100 μm. See also Figures S3 and S4 and Table S1. Cell Reports 2017 21, 3653-3661DOI: (10.1016/j.celrep.2017.12.026) Copyright © 2017 The Authors Terms and Conditions