Figure 5. Comparative NBE/TPE proteomics. Serum levels of 308 mouse proteins (Raybiotech #L-308) and 507 human proteins (Raybiotech #L-507) were assayed. After background subtraction, total intensities for each protein (assayed in duplicates for each sample) were normalized to the internal array background control as fold-increments, with a sensitivity cut-off of 2-fold; these normalized intensities were expressed, as a fraction of the internal array positive control. (A) t-SNE clustering of mouse proteins grouped by class of treatment: OO and YY isochronic controls were compared to each other (left) and OO was compared to ONBE (right). Differences between YY vs. OO and OO vs. ONBE proteomes are outlined. (B) Distinct grouping of OO, YY and ONBE proteomes is shown in the t-SNE plot with identities of proteins in clusters 1-4 specified below. Power analysis for independence of X, X and Y marked proteins from these clusters, is shown in Supplementary Figure 4. (C) Heatmap on mouse proteins illustrates significant differences between OO and ONBE cohorts (proteins are grouped on their main function, as indicated). (D) Old human serum proteome before TPE (Before-B) and 1 month after a single procedure of TPE (After-A): t-SNE clustering of human proteins grouped by class of treatment. TPE resulted in a clear and robust change in the molecular composition of the systemic milieu as compared to the Before-TPE. (E) Heat map on human proteins illustrates significant differences between S1,2,3 B and S1, 2, 3 A (before versus after TPE) cohorts (proteins are grouped on their main function, as indicated). Proteins in dashed boxes are the same between mouse and human in Heatmaps. A general elevation (not decrease) of most systemic proteins at 6 days after the NBE and at 1 month after the TPE was observed.