Skip to main content
. Author manuscript; available in PMC: 2022 Jul 18.
Published in final edited form as: Nat Cell Biol. 2021 Jun 9;23(6):631–641. doi: 10.1038/s41556-021-00693-y

Extended Data Fig. 10. Analysis of ubiquitous and abundant exosomal proteins using different approaches.

Extended Data Fig. 10

(a) Western blot analysis of ITGB1, LGALS3BP, SLC3A2, Alix, CD47 and TSG101 in UC-isolated exosomes from HEK293T cells. Results from one biological replicate. (b) Western blot analysis of ITGB1 in exosomes from the OptiPrep fractions of HEK293T, MDAMB231 and PANC-1. Results from one biological replicate. (c) Western blot analysis of ITGB1 in exosomes from the exosome-rich and exosome-depleted SEC fractions, from HEK293T, MDAMB231 and PANC-1. Representative results from three biological replicates. (d) Representative histograms show CD47 and ITGB1 levels in comparison to isotype control-stained beads in HEK293T, MDAMB231 and PANC-1-derived exosomes. Bar graph shows mean +/− s.e.m. of percentage of positive beads, individual data points from three biological replicates. (e) Representative histograms show the profile of the CD47 and ITGB1 levels in comparison to isotype control-stained exosomes from human plasma. Bar graph shows MFI +/− s.e.m, and individual data points refer to ten donors. Statistical significance was determined using two-tailed unpaired Mann Whitney t-test, and the exact p-values are shown. Significance defined as p < 0.05. (f) Gating strategy used for the FACS-based single particle analysis of exosomes. (g) Representative histograms show Syntenin-1 and CD63 positivity in non-permeabilized and permeabilized exosomes, in comparison to isotype control stained beads-bound exosomes.