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. 2021 Oct 14;10(12):e12158. doi: 10.1002/jev2.12158

FIGURE 5.

FIGURE 5

Effect of urine storage temperature (‐20°C vs. ‐80°C) on uEV morphology and protein markers. a. Transmission electron micrographs of frozen uEV from donors with type 1 diabetes and normo‐ or macroalbuminuria. Urine samples were stored at ‐20°C or ‐80°C for 1 year before uEV isolation. b. Western blotting of uEV isolated from urine stored at ‐20°C or ‐80°C for 1.5 months (healthy) or longer, up to 4 years (normo and macro). Here, uEV were isolated from low urine volume, 7.8 ml (uEV isolated from high urine volumes in Supplementary Figure 3). uEV derived from equal volumes of urine were loaded for each sample. Immunodetection of EV markers (PDX, TSG101, CD9, CD63 and CD59), a common protein that co‐isolates with uEV (THP) and a Golgi marker (GM130). Healthy control (Healthy), macroalbuminuria (macro), normoalbuminuria (normo), podocalyxin (PDX), Tamm–Horsfall protein (THP), urinary extracellular vesicles (uEV)