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. 2016 Nov 3;22(11):729–744. doi: 10.1093/molehr/gaw049

Figure 5.

Figure 5

CRES subgroup proteins are associated with extracellular vesicles in the mouse epididymal luminal fluid. Extracellular vesicles were isolated from initial segment/caput epididymal luminal fluid (LF) (Segments 1–5) from Cst8(+/+) and Cst8(–/–) mice using the Exoquick reagent and resuspended in RIPA buffer (50 mM Tris, pH 7.4, 150 mM NaCl, 1% Triton X-100, 0.1% SDS, 0.5% sodium deoxycholat, 1 mM EDTA). Seventy micrograms of protein were loaded onto SDS-PAGE gels and immunoblot analysis carried out using anti-CRES, anti-CRES2, anti-CRES3, anti-cystatin E2 and anti-CD9 antibodies. About 20 µg LF (prior to isolation of vesicles) from the initial segment/caput epididymis (Segments 1–5) served as a positive control.