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. 2014 Aug 30;17(1):105–118. doi: 10.1111/cmi.12339

Figure 2.

figure

Identification of IFITM3‐containing exosomes derived from 293T and HUVEC cells.

A–D. Cell culture supernatants of HUVEC or HepG2 cells transfected with IFITM3 siRNA‐1 (HUVEC‐siIFITM3 or HepG2‐siIFITM3), from HUVEC or HepG2 transfected with non‐targeting control siRNA (HUVEC‐siNC or HepG2‐siNC), from293T cells transfected with pcDNA3 (293T‐Vector), or from 293T cells transfected with the pcDNA‐IFITM1, 2, 3‐Flag construct (293T‐IFITM1, 2, 3) for 48 h, were collected and concentrated for SDS‐PAGE and immunoblotting, using an anti‐IFITM3 antibody, with cell lysates as positive control.

E. Electron micrographs of crude exosomes negatively stained with uranyl acetate and examined at 80 kV are shown.

F. Purified IFITM3‐containing exosomes derived from each group of cells above described were analysed by immunoblotting with anti‐IFITM3, anti‐flotillin‐2, anti‐CD63, anti‐calnexin (endoplasmic reticulum, ER marker) and anti‐GM130 (Golgi marker) antibodies. IFITM3 is identified in the exosomes derived from each group of cells as indicated.