Enterocloster bolteae drives hyperglycemia-associated colorectal cancer cell proliferation via Rac/Rho-MSH6-Elongin C signaling pathway

Figure 5 Functional analysis based on the Kyoto Encyclopedia of Genes and Genomes database. A: Statistical chart of the number of annotated genes in each database. The numbers on the bar chart represent the number of unigenes successfully annotated. The other axis represents the level 1 functional categories within each database; B: Sample clustering analysis based on functional abundance. The left side of the figure displays the clustering tree structure based on Bray-Curtis distance. The right side shows the relative abundance distribution of functions at level 1 for each sample; C: Functional abundance clustering heatmap based on the top 5 most abundant functions per sample, using the functional annotations and abundance data from the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. The horizontal axis represents sample information, and the vertical axis provides functional annotation information. The clustering tree on the left represents functional clustering. The values in the central heatmap correspond to the Z-scores obtained by normalizing the relative abundance of functions in each row; D: Top 10 KEGG level 1 functional ranked by relative abundance across samples. The Y-axis indicates the proportion of annotations within each functional category, and the X-axis shows the sample names. Relative abundance was calculated at multiple levels across databases. This figure shows the relative abundance distribution at KEGG level 1 as an example; E-G: Dimensionality reduction analysis based on functional abundance in the KEGG database. The more similar the functional composition between samples, the closer their distances in the dimensionality reduction plot. When the stress value of NMDS analysis at level 1 is less than 0.2, the non-metric multidimensional scaling analysis is considered to have a certain degree of reliability (Figure 5E). Principal component analysis at level 1 is shown, where the horizontal axis represents the first principal component, the vertical axis represents the second principal component, and the percentages represent the contribution of each component to sample differences (Figure 5F). Dimensionality reduction analysis based on Bray-Curtis distance derived from species abundance was also performed, as illustrated by principal coordinate analysis at level 1 (Figure 5G). DM: Diabetes mellitus; CRC: Colorectal cancer; PCA: Principal component analysis; NMDS: Non-metric multidimensional scaling; PCoA: Principal coordinate analysis.