Extended Data Fig. 10: PFNA does not affect gut microbiota composition of mice colonised with human gut bacteria.

a. Gut microbiota composition of mice colonised with 20 human gut bacterial strains (Com20) and exposed to 10 mg/kg body weight by oral gavage on day 0. 17 out of 20 gut bacteria colonised the mice with 7 strains making up 90% of bacterial composition (P. vulgatus, R. gnavus, B. thetaiotaomicron, B. uniformis, A. rectalis, Veillonella parvula, Dorea formicigenerans). The composition in the small intestine differed from that in the feces and colon, with the dominating species being R. gnavus and V. parvula (Supplementary Table 42). b. In the colon and fecal samples 70% of bacteria were classified as high-PFNA accumulating strains, while in the small intestine only 20% of bacteria were classified as high-PFNA accumulating (Supplementary Table 43). c. Gut microbiota composition of mice colonised with a community of five high-PFNA accumulating strains (HC) and five low-PFNA accumulating strains (LC) exposed to 10 mg/kg body weight by oral gavage on day 0. All gut bacteria colonised the mice, but A. muciniphila was present in both HC and LC colonised mice (Supplementary Table 44). d. LC (low-PFNA accumulating community) colonised mice showed higher DNA yields after DNA extraction compared to HC (high-PFNA accumulating community) colonised mice (1.96-fold change, p-value = 0.002), indicating higher bacterial content in LC colonised mice; two-sided t-test; Boxplot: centre = 50th percentile, bounds of box = 25th and 75th percentile, lower/upper whisker = lower/upper hinge −/+ 1.5 * inter quartile range; n = 54 samples per group (Supplementary Table 28). e. HC colonised mice showed higher relative abundance of Acetobacter fabarum spike-in compared to LC colonised mice (2-fold change, p-value = 0.0009), indicating higher bacterial content in LC colonised mice. Two-sided t-test; Boxplot: centre = 50th percentile, bounds of box = 25th and 75th percentile, lower/upper whisker = lower/upper hinge −/+ 1.5 * inter quartile range; Subset of 76 samples (Supplementary Table 45). f. Mice colonised with a community of 20 human gut bacterial strains (Com20) show higher PFNA excretion after 0.1 mg/kg body weight PFNA exposure compared to germfree (GF) controls; Boxplot: centre = 50th percentile, bounds of box = 25th and 75th percentile, lower/upper whisker = lower/upper hinge −/+ 1.5 * inter quartile range; two-sided t-test; p-values are FDR corrected; n = 6 mice per group; y-axes are on log₁₀ scale (Supplementary Table 46).