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Figure 2. Differential protein expression and functional analyses of APS-Lat-EXO and Lat-EXO. (A) Venn diagram showing uniquely and commonly identified proteins in APS-Lat-EXO and Lat-EXO; (B) Volcano plot of differentially expressed proteins, with FC shown on the log₂ scale (x-axis) and statistical significance expressed as -log₁₀ P values (y-axis). Proteins with increased abundance are highlighted in red, while those with decreased abundance are shown in green (|log₂FC| ≥ 1.00; P ≤ 0.01); (C) Hierarchical clustering heatmap of protein expression profiles in APS-Lat-EXO and Lat-EXO, with three biological replicates per group; (D) PCA score plot showing the distribution of samples along the first two principal components; (E) PLS-DA score plot based on 758 differentially expressed proteins screened using Ranking-PCA; (F-H) Expression levels of MKI67, CD38, and CXCR4 in exosomes; (I-K) GO enrichment analysis of the 758 differentially expressed proteins, showing the top 20 categories in CC, biological process, and molecular function; (L) KEGG pathway enrichment analysis of the same protein set, showing the top 20 significantly enriched pathways; (M) Representative subsets of differentially expressed proteins associated with PI3K/AKT signaling, actin cytoskeleton regulation, endocytosis, focal adhesion, Rap1 signaling, HIV-1 infection, and leukocyte transendothelial migration; (N) Expression levels of AKT1, AKT2, and AKT3 in exosomes; (O) Cytoscape network map of enriched GO terms; (P) Cytoscape network map of enriched KEGG pathways. Statistical analysis was performed using an unpaired t test with Welch’s correction (n = 3). Data are presented as mean ± SD. ^*P < 0.05; ^**P < 0.01; ^***P < 0.001. APS: Astragalus polysaccharide; APS-Lat-EXO: APS-modified latency-reversing exosome; log₂FC: log₂ fold change; PCA: principal component analysis; PLS-DA: partial least squares discriminant analysis; MKI67: marker of proliferation Ki-67; CD38: cluster of differentiation 38; CXCR4: C-X-C chemokine receptor type 4; GO: Gene Ontology; KEGG: Kyoto Encyclopedia of Genes and Genomes; PI3K: phosphoinositide 3-kinase; AKT: protein kinase B; SD: standard deviation.