Western Blotting of Extracellular Vesicle Proteins

Western blotting was performed as described previously with modifications [32 (link)]. Cells or EVs were lysed in 1x RIPA buffer with protease inhibitors. The lysate was centrifuged and the supernatants collected. Mouse monoclonal anti-beta-actin (Sigma), mouse monoclonal anti-C3b (Thermo scientific), mouse monoclonal anti-properdin (Abcam), rabbit monoclonal anti-HSP70 (Abcam), rabbit polyclonal anti-CD59 (Abcam), rabbit polyclonal anti-CD55 (Santa Cruz Biotechnology), mouse monoclonal anti-CD63 (Santa Cruz Biotechnology), mouse monoclonal anti-CD81 (Santa Cruz Biotechnology), Rabbit polyclonal anti-CD46 (Santa Cruz Biotechnology), Rabbit polyclonal anti-Calnexin (Bioss Antibodies Inc.), and rabbit polyclonal anti-CD55 (Santa Cruz Biotechnology) were used.

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Jeon H., Yoo S.M., Choi H.S., Mun J.Y., Kang H.G., Lee J., Park J., Gao S.J, & Lee M.S. (2017). Extracellular vesicles from KSHV-infected endothelial cells activate the complement system. Oncotarget, 8(59), 99841-99860.

Publication 2017

Mouse monoclonal anti-beta-actin mouse monoclonal anti-C3b mouse monoclonal anti-properdin rabbit polyclonal anti-CD59 rabbit polyclonal anti-CD55 mouse monoclonal anti-CD63 mouse monoclonal anti-CD81 Rabbit polyclonal anti-CD46 Rabbit polyclonal anti-Calnexin

Antibodies Beta actin Buffer Calnexin Cd59 Cells Hsp70 Mouse Properdin Protease inhibitors Rabbit Ripa

Corresponding Organization : Eulji University

Other organizations : University of Southern California

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Variable analysis

independent variables

Cell or EV lysis in 1x RIPA buffer with protease inhibitors

dependent variables

Expression levels of beta-actin, C3b, properdin, HSP70, CD59, CD55, CD63, CD81, CD46, Calnexin

control variables

Not explicitly mentioned

controls

No positive or negative controls were explicitly mentioned.

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