Figure 2
Extracellular Vesicles are released from CHLA 02 and CHLA 04 cells and contain HML-2 Envelope (Env). (a,b) AT/RT cells release extracellular vesicles contain HML-2 Env. (a) Immunostaining CHLA 02 cells with HML-2 env polyclonal antibody (surface unit) (red) and CD98 (green), a cell-surface marker, shows Env is expressed in extracellular vesicles enclosed by the plasma membrane. Cell nuclei are stained with DAPI (blue); (b) CHLA 04 cells similarly immunostained; magnification is at 63X. (c) CHLA 02 cells incubated with secondary antibody only as a control for nonspecific immunostaining. (d,e) Electron microscopy shows extracellular vesicles forming on plasma membrane of CHLA 02 AT/RT cells (arrow heads). (f) Cell lysates from Tera, CHLA 02, and CHLA 04 cells express HML-2 env (left three lanes) while only purified extracellular vesicles from CHLA 02 and CHLA 04 express HML-2 env (three right lanes) on immunoblot (arrow). A fluorescent Zeiss confocal microscope (LSM510) was used to acquire images processed with Zeiss LSM 5 Image Browser and Microsoft PowerPoint for figure creation (https://www.embl.de/eamnet/html/downloads.html). The gel image has been cropped to focus on the bands of interest, the full gel image can be found in the supplementary material. Gel Image was taken with a FluorChem E system (https://www.proteinsimple.com/fluorchem.html) and images were processed with ImageJ and Microsoft PowerPoint for relative densitometry quantification and display. Electron microscopy images were taken with a JEOL 1200 EXII Transmission Electron Microscope (AMT digital camera system).
