Extended Data Fig. 6: Mucin utilization is required for A. muciniphila to compete in CONV mice and in Muc2-/- mice.
A breeding colony of Akkermansia-free mice (Akk-free) was generated to facilitate mouse colonization without antibiotic pre-treatment. (a-c) Comparison of the microbiota of Akkermansia colonized (Akk-colonized) and Akk-free mice by 16 S rRNA gene sequencing. (a) Relative abundances of fecal bacteria at the genus level in Akk-colonized and Akk-free mice. Each sample was obtained from separately housed mice (n = 3 per group). (b) Principal Coordinates Analysis (PCoA) performed on weighted UniFrac distances. Statistical significance was determined by Permutational Multivariate Analysis of Variance (PERMANOVA). (c) Relative abundances of potential mucin-degrading taxa at the family level. The centre line is the mean, and the whiskers show the minimum and maximum. (d) Linear discriminant analysis Effect Size (LEfSe) analysis of Akk-colonized and Akk-free mice. The Kruskal-Wallis test was used to detect features with a significant differential abundance (p < 0.05). (e) CONV mice were pre-treated with antibiotics and gavaged with a 1:1 mix of WT and mutant A. muciniphila prepared with a fecal slurry from Akk-free mice to partially reconstitute the microbiota (n = 6 per group). Bacterial loads in fecal pellets were quantified by qPCR, each point represents one cage. (f) Colonization of mucin deficient Muc2-/- mice with A. muciniphila. Each point represents the average A. muciniphila per gram of feces (WT, n = 4; mul1A::Tn, n = 4; mul2A::Tn, n = 6). (g) Competition between wild type A. muciniphila and the mul1A::Tn mutant in Muc2-/- mice. Mice were gavaged with a 1:1 mix of wild type and mutant and abundance was monitored over time using strain specific primers. Each point represents the average amount of A. muciniphila (n = 4), error bars represent the standard error.
