Fig. 6

Microbial functional alterations in irAEs and non-irAEs. A Differentially abundant pathways were plotted; P values were computed using a two-sided blocked Wilcoxon rank-sum test and the FDR < 0.005 were presented in the heatmap. B The correlation between the abundance of menaquinone biosynthesis-related pathway and model species. Spearman’s correlation between the abundance of menaquinone biosynthesis and 14 representative microbial species in the classifiers and edge width corresponds to the Spearman’s r statistic and edge color denotes the statistical significance. r, Spearman correlation coefficient; with a color-gradient denoting Spearman’s correlation coefficients, and the exact values were described in heatmap frames. C, D Plotted values are qRT-PCR quantifications of bacterial genes in menaquinone biosynthesis. The abundances of menC (C) and menH (D) were compared between non-irAEs (n = 17) and irAEs (n = 11) groups. All boxes extend from 25 to 75th percentiles and whiskers show the minimum and maximum values. Lines at the middle of each box show the median. P values were computed using a two-sided Wilcoxon rank-sum test. E–M Shotgun metagenome functional validation (N = 50) for differential genes menC (E), menH (F), and menaquinone biosynthesis pathway (G–M). All boxes extend from 25 to 75th percentiles and whiskers show the minimum and maximum values. Lines at the middle of each box show the median. P values were computed using a two-sided Wilcoxon rank-sum test. O Blood concentration comparison of menaquinone-6 (MK-6) between patients with irAEs( N = 10) and patients without irAEs(N = 10). Statistical significance was assessed using a two-sided Wilcoxon rank-sum test