Fig. 2

Proteomic profile of CHMp and CHMm-derived exosomes. A Schematic figure of exosome proteomics analysis. B TMT labeling intensity efficiency between CHMp and CHMm exosomal proteins in triplicates. C-D Principal Component Analysis (PCA) of CHMp and CHMm exosomal proteome replicates. C TMT intensity correlation of the replicates of CHMp and CHMm exosomal proteins. PCA showed high correlation for technical and biological replicates. D PCA showed high correlations of CHMp and CHMm exosomal protein triplicates. The X and Y axes show principal component 1 and principal component 2, respectively. E Heatmap of 13 exosomal proteins representing cellular localization. CD9, CD63, CD82, TSG101, Alix and GAPDH for exosome, HSP90B1, HIST2H3A and SUB1 for nucleus, MAN2A1, BTGALT1 for golgi, LAMP1 for lysosome, ITGB5 for mitochondria. F Volcano plot based on the Log2 (fold change) and their -Log10 (p-value) of CHMp and CHMm exosomal proteins. In the plot, red dots indicate the proteins that are statistically enriched in CHMp exosomes, blue dots represent proteins enriched in CHMm exosomes, and grey dots represent proteins that are not statistically significant. Significantly enriched proteins in CHMp exosomes compared with CHMm exosomes as control, Student’s t-test, p < 0.05, obtained in Perseus software. G The subcellular localization of CHMp and CHMm exosomal proteins showing significantly differential expressions. Predicted subcellular localization were obtained from UniProt (https://www.uniprot.org/). CHMp exosomal proteins were predominantly located in the nucleus and extracellular region. n = 3 biologically independent exosomal protein isolations. Figure 2A created with BioRender.com