Fig. 5.

Subcellular localisation and sorting to sEV of UBLs. a Representative images of MDA-MB-231 cells transfected either with EGFP-ubiquitin, -SUMO1, -SUMO2 or -UBL3 and co-stained with markers for MVBs (CD63). Scale bars, 10 and 1 μm. b Quantitative analysis of EGFP-ubiquitin, -SUMO1, -SUMO2, and -UBL3 fluorescence intensity in a. EGFP-ubiquitin, n = 10; EGFP-SUMO1, n = 10; EGFP-SUMO2, n = 10; EGFP-UBL3, n = 10; EGFP, n = 10. *, p < 0.05; ***, p < 0.0001 by Kruskal–Wallis/Dunn's multiple-comparisons test. c EGFP-UBL3, but not EGFP, EGFP-ubiquitin, EGFP-SUMO1, or EGFP-SUMO2, was preferentially enriched in the sEVs of the cell culture media. Before sample loading, the samples were boiled with βME