fig3

Figure 3. Differential miRNA expression and functional analyses of APS-Lat-EXO and Lat-EXO. (A) PCA plot based on exosomal miRNA expression profiles; (B) Hierarchical clustering heatmap of differentially expressed miRNAs, with three biological replicates per group; (C) Volcano plot depicting miRNA expression changes between APS-Lat-EXO and Lat-EXO; log₂ fold change is plotted on the horizontal axis and -log₁₀ P value on the vertical axis. Upregulated miRNAs are highlighted in red, whereas downregulated miRNAs are shown in blue (P < 0.05, FC > 5); (D-G) Relative expression levels of the four miRNAs that were downregulated (hsa-miR-92a-3p, hsa-miR-8075, hsa-miR-297, and hsa-miR-1290); (H-J) Gene Ontology enrichment results of predicted target genes associated with the four downregulated miRNAs; (K) KEGG pathway enrichment analysis of the corresponding target genes; (L) Cytoscape-based network visualization of miRNA target genes annotated by GO terms; (M) Cytoscape network representation of miRNA target genes mapped to KEGG pathways; (N) Cytoscape network map of predicted TF-miRNA interactions. Statistical comparisons were performed using an unpaired t test with Welch’s correction (n = 3). Data are presented as mean ± SD. ^**P < 0.01. APS: Astragalus polysaccharide; APS-Lat-EXO: APS-modified latency-reversing exosome; PCA: principal component analysis; FC: fold change; GO: Gene Ontology; KEGG: Kyoto Encyclopedia of Genes and Genomes; TF: transcription factor; SD: standard deviation.