Skip to main content
. 2019 Jun 4;10(38):3694–3708. doi: 10.18632/oncotarget.26979

Figure 1. MAGEA proteins are incorporated into EVs.

Figure 1

(A) Schematic representation of the purification scheme of EVs. (B) Western blot analysis of COP5 cells and EVs purified from COP5 cells ectopically expressing recombinant MAGEA proteins. The presence of proteins was analysed by specific antibodies against MAGEA4, MAGEA10, TSG101 (T5701; Sigma-Aldrich) and alpha-tubulin (T5168; Sigma-Aldrich). 5% of each EV fraction was loaded to the gel. (C) Western blot analysis of EVs purified from COP5 cells ectopically expressing recombinant MAGEA proteins using anti-E2Tag antibody (for MAGEA4 and MAGEA10) and anti-TSG101 antibody (T5701; Sigma-Aldrich). 5% of each EV fraction was loaded to the gel. (D) Western blot analysis of EVs obtained with ultracentrifugation through stepwise sucrose density gradient (20%, 35%, 45%, 60%) at 120 000 g for 1.5 hours at 4° C. Gradient was divided into 8 fractions and the presence of MAGEA4 protein in each fraction was analyzed with MAGEA4 specific antibody. Top and bottom on the right size of the image depicts the loading place of samples. (E) Physical characterization of EVs as assessed by DLS (Dynamic Light Scattering). The hydrodynamic diameter (Z-average) and polydispersity index (PdI) are shown on the right. (F) Physical characterization of 120K EVs as assessed by NTA (Nanoparticle tracking analysis).