Fig. 5: Entacapone-exposed microbiota cells display higher iron content.

a,b, NanoSIMS overlay images of the ⁵⁶Fe⁺ (colour scale) and ¹²C⁺ (grey scale) signal intensities of a faecal sample incubated for 6 h without drug (a) or ENT-Hi (b) in sM9 medium. Scale bars, 10 μm. c, Evaluation of ⁵⁶Fe⁺/¹²C⁺ secondary ion signal intensity ratios in individual cells present in faecal samples. d,e, NanoSIMS overlay images of the ⁵⁶Fe⁺ (colour scale) and ¹²C⁺ (grey scale) signal intensities in P. dorei cells incubated for 24 h with no drug (d) or ENT-Hi (e) in BMM medium. In the individual ¹²C⁺ images used for merging, the minimum and maximum intensities (grey scale) are: 1–16 counts per pixel (a), 2–25 counts per pixel (b), 4–80 counts per pixel (d) and 0–25 counts per pixel (e). In the individual ⁵⁶Fe⁺ images used for merging, the minimum and maximum intensities (colour scale) are: 3–10 counts per pixel (a), 3–35 counts/ per pixel (b), 7–17 counts per pixel (d) and 4–80 counts per pixel (e). Scale bar, 10 μm. f, Evaluation of ⁵⁶Fe⁺/¹²C⁺ intensity ratios in individual P. dorei cells. Levels of significance (P values) displayed in c and f were calculated using unpaired two-sided Wilcoxon test comparing the groups ‘Ent-Hi’ and ‘No drug’. Each dot represents a single cell and boxes represent the median, first and third quartiles. Whiskers extend to the highest and lowest values that are within 1.5× the interquartile range. Single-cell measurements and calculations are shown in Supplementary Table 18.