Fig. 3: Analyses of the proteomes of BA9 EVs from control and SUD BA9 brain tissues.

A, B Hierarchical cluster analysis assessing differences in BA9 EVs from control and SUD BA9 tissues using Pearson’s correlation reveals differences visualized as A heatmaps and B correlations. C Volcano plot analysis of the BA9 proteome. D Gene Ontology analysis showing the abundance of proteins originating from exosomes, vesicles, and the extracellular space, confirming successful enrichment of vesicles derived from brain tissue. E Gene Ontology analysis of the BA9 proteome using the WEB-based GEne SeT AnaLysis Toolkit (WebGestalt 2024). F Ingenuity pathway analysis (IPA) was used to predict ontologically related functions and molecules predicted to be increased (orange) or decreased (blue) when driven by SUD-induced changes in the proteome of BA9 EVs. The DEGs are involved in biological processes such as cell viability, cell migration, and degranulation and are not involved in disease, while the genes encoding IL1B, AGT, STAT3 and others may regulate these functions. A solid line represents a direct interaction between two gene products, and a dotted line indicates an indirect interaction. G IPA-identified upstream regulators of cellular functions related to cell death (left) and cell migration (right) mediated by BA9 EVs. A solid line represents a direct interaction between two gene products, and a dotted line indicates an indirect interaction. Orange indicates activation or increase, and blue indicates inhibition or decrease. H IPA-identified most highly rated network and canonical pathway analysis. I IPA-identified groups of ontologically related diseases predicted to be linked to the proteome of BA9 EVs.