Fig. 3: TMEM33-marked R-EVs are distinct from classical exosomes.
From: The assembly of RAB22A/TMEM33/RTN4 initiates a secretory ER-phagy pathway
a Schematic diagram depicting the workflow for the immunoaffinity approach for isolating FLAG-tagged R-EVs and CD63-positive classical exosomes from total EVPs collected from HeLa cells stably expressing both GFP-RAB22AQ64L and FLAG-TMEM33. b The cell lysates, total EVPs and anti-FLAG or anti-CD63 bead-conjugated EV populations were analyzed via western blotting using the indicated antibodies. c The purified FLAG-TMEM33-labeled R-EVs eluted with FLAG peptides were attached to coverslips precoated with 0.2 mg/mL poly-L-lysine. Then the R-EVs were stained with the indicated antibodies. Scale bar, 1 μm.
