Integrative Metabolomic and Proteomic Analysis

The metabolomics and proteomics data, after a batch-effect adjustment and log transformation, were analysed using multivariate data analysis software SIMCA (version 16, Sartorius Stedim Biotech, Umeå, Sweden) and MetaboAnalyst 4.0 [29 (link)]. The Gene Ontology Resource and Enrichr were used for Enrichment analysis. The cut-off level for significant metabolites was a signal-to-noise (S/N) ratio of 10, while for proteins, it was a relative abundance of 1 × 10⁵. For statistical analysis of both metabolome and proteome, a fold change of ≤0.5 (downregulation) or ≥2.0 (upregulation), and a Benjamini–Hochberg adjusted p-value of ≤0.05 was used. Metabolomic and proteomic outputs were integrated using the ‘Joint-pathway analysis tool’ of MetaboAnalyst 4.0 and Paintomics 3 [30 (link)], and the false discovery rate (FDR) was used to report adjusted p-values.