General descriptive statistics of sample population

Top ten differentially expressed Proteins between normal and preeclampsia placentas (bold font), medium/large STB-EVs (normal font), small STB-EVs(italics)

Flow analysis of medium/large STB-EVs in the 10K STB-EV pellet. Apogee beads mix were used to set the flow machine’s light scatter resolution to 0.59-1.3 mm and 0.24-1.3

(A) mm silica beads. Figures B show the application of SSC and FSC PMTVs as determined by apogee beads mix for the analysis of m/lSTB-EVs in the 10K pellet. An investigation gate was created to include only medium/large EVs negative for non-placental markers (C & F). Extracellular vesicles from the investigation gate were further analyzed for staining by bioM and expression of PLAP (D & G). Figure E and H shows that the bioM+ PLAP+ EVs were sensitive to detergent treatment. The percent of B-Maleimide+ PLAP+ from the 0.59-1.3 micron gate is consistent under both SSC conditions.

Results of STB-EV characterization. Figure 2A and 2D show the NTA results of S STB-EVs (2D) and m/lSTB-EVs (2A). Figure 2B,2C,2E and 2F displays representative transmission electron microscopy (TEM) images with wide view (2B and 2E), medium/large STB-EVs (2C), and Small STB-EVs (2F).

List of overlapping differentially carried proteins in the placenta, medium/large STB-EVs and small STB-EVs

Flow chart illustrating the steps involved in characterizing m/lSTB-EVs and sSTB-EVs obtained via differential ultracentrifugation (10,000 and 150,000 g) of maternal perfusate. Nanoparticle trafficking analysis, transmission electron microscopy, flow cytometry, and western blot were used to characterize the pellets from both spins.

Reagents used for western blot.

Antibodies used for western blot.

The top three functionally enriched gene ontologies are: biological process (GO: BP) Placenta (bold font), medium/large STB-EVs (normal font), and small STB-EVs (italics).

The top three functionally enriched KEGG Pathways in the placenta, medium/large STB-EVs, and small STB-EVs (only 1 KEGG). Placenta (bold font), medium/large STB-EVs (normal font), and small STB-EVs (italics)