Skip to main content
. Author manuscript; available in PMC: 2020 Aug 6.
Published in final edited form as: Cell Metab. 2019 Jun 13;30(2):329–342.e5. doi: 10.1016/j.cmet.2019.05.015

Figure 6. Plasma eNAMPT is exclusively localized to EVs in both mice and humans.

Figure 6.

(A) Comparison of eNAMPT, EV marker proteins (TSG101, CD63, CD81, and CD9), and non-EV proteins (transferrin and albumin) in whole plasma, EV fraction, and supernatant isolated by ultracentrifugation and the Total Exosome Isolation (TEI) kit. The protein concentrations were typically ~0.4 and ~1 μg/μl for EVs purified by ultracentrifugation and the TEI kit, respectively, when EVs were reconstituted with an equal volume of PBS to the starting volume of plasma. 40 μg of protein from each fraction were loaded.

(B) Comparison of eNAMPT and EV marker proteins in six fractions (F1-6) isolated from sucrose density-gradient centrifugation. 2 ml of plasma were used for this fractionation.

(C) Comparison of eNAMPT in whole plasma (P), EV fraction (E), and supernatant (S) isolated from three 4 month-old male mice and 37, 41, and 45 year-old male human donors. Each fraction was loaded after adjusting them to an equal volume.

(D) Comparison of eNAMPT, TSG101, transferrin, and immunoglobulin light chain (Ig LC) in the treatment of mouse plasma with proteinase K and/or Triton X-100.

(E) Levels of EV-contained eNAMPT (EV-eNAMPT) and CD63 in the plasma from 6 and 22 month-old mice (n=4 per group).

(F) Levels of EV-contained eNAMPT (EV-eNAMPT) and CD63 in the plasma of control (CTRL) and ANKI mice at 24 month of age (n=4 per group).