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. 2021 Jan 19;22(2):979. doi: 10.3390/ijms22020979

Figure 5.

Figure 5

Protein concentration and number of proteins identified in equine uterine extracellular vesicles (EVs). (A) Comparison of protein concentration (µg/µL) of EVs isolated by different methods. (B) Comparison of the number of proteins identified in EVs by mass spectrometry among methods. Different letters indicate significant differences (a,b: p < 0.05). EVs isolation methods: MA_UC: EVs obtained by ultracentrifugation (UC) using Beckman fixed angle rotor from 10, 20, and 40 mL of uterine flush, and from 10 mL PBS as a control; MB_UC: EVs obtained by Beckman swing-out rotor; MC_CE/PBS: EVs obtained by ultrafiltration using Centricon filters (CE), wash step with PBS and followed by UC swing-out rotor; MD_CE/TRE: similar to before but wash step with PBS/25 mM trehalose; ME_CE/IZON/UC: EVs obtained by combination of CE, followed by SEC, Izon qEV column and collection of fractions enriched in EVs (7,8,9) and UC swing-out rotor; and MF (MF_CE/TRE_IZON/UC): CE with PBS/25 mM trehalose wash step, Izon qEV columns and UC.