Skip to main content
. Author manuscript; available in PMC: 2020 Apr 4.
Published in final edited form as: Cell. 2019 Apr 4;177(2):428–445.e18. doi: 10.1016/j.cell.2019.02.029

Figure 1. High-Resolution Density Gradient Fractionation Separates Small Extracellular Vesicles from Non-Vesicular Components.

Figure 1.

(A) Nomenclature of extracellular vesicles and particles employed in this study.

(B) Immunoblots of DKO-1 and Gli36 whole cell lysates, large EVs (P15) and crude small EVs (P120) (STAR Methods). Equal quantities of protein were separated on SDS-PAGE gels, and membranes were blotted with indicated antibodies.

(C) Density gradient fractionation of DKO-1 and Gli36 crude small EVs (P120). After flotation of sample in high-resolution iodixanol gradients (STAR Methods), equal volumes of each fraction were loaded on SDS-PAGE gels, and membranes were blotted with indicated antibodies. NV, non-vesicular; sEV, small EV.

(D) Nanoparticle tracking analysis of pooled low (sEV) and high (non-vesicular) density fractions with or without pre-treatment with Triton X-100 detergent. For box plots the center lines mark the median; box limits indicate 25th and 75th percentiles; whiskers extend 1.5 times the interquartile range from 25th and 75th percentiles; n = 18 sample points; data from three independent experiments. Significant differences were assessed by one-way ANOVA and pairwise comparisons adjusted by the Holm-Bonferroni method; *p < 0.001; N.S., Not Significant.

(E) Negative stain transmission electron microscopy (TEM) of DKO-1 large EVs (P15), and pooled low (sEV) and high (NV) fractions obtained from high-resolution density gradients.

(F) Negative stain TEM of Gli36 pooled low (sEV) and high (NV) fractions. See also Figure S1 and Table S1.