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. 2021 Apr 15;13(4):675. doi: 10.3390/v13040675

Figure 6.

Figure 6

The absence of CD63 leads to increased interaction of mTOR, Nedd4L, PP2A. (A) Both Nedd4L and PP2A delta interact with CD63. Pull-down assays using CD63 BioID fusion proteins Figure 2. A delta with CD63. (B,C) Immunoprecipitation of mTOR, Nedd4L, and PP2A in #1 B-cells. Immunoprecipitation in the #1 cells was carried out using LMP1 antibodies and CD63 pre-conjugated beads followed by immunoblotting. (D) Pull-down experiments using LMP1 BioID fusion proteins followed by Western blotting using indicated antibodies demonstrates that LMP1 BioID fusion proteins pulled-down an increased level of the interacting proteins in the absence of CD63. At the same time, WCE shows a similar level of proteins in all the lanes. (E) The increased interaction with LMP1 in absence of CD63 is not due to differential targeting of the extracellular vesicles (EVs). EVs purified from supernatant were resolved on SDS-PAGE and blotted for the antibodies indicated. The result indicates that there is no obvious difference in EV targeting due to the absence of CD63.