Supplementary Figure 1 a b c i ii iii i ii iii i ii iii

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Supplementary Figure 1 a b c i ii iii i ii iii i ii iii Tg rat (E18-19) Tg rat (E18-19) i ii iii i ii iii i ii iii GFP- GFP++ GFP+ M 1 2 3 4 5 6 7 1 2 3 4 5 6 7 8 9 CAG/CD63-GFP BF GFP Merged i: GFP- ii: GFP++ iii: GFP+ Sry GFP- GFP++, + GFP++ GFP+ 1 2 3 4 5 6 7 1 7 1 2 3 4 5 6 7 8 9 rat CD63 BF GFP Merged BF GFP Merged BF GFP Merged human CD63 copGFP β-actin c Tg rat (E17) GFP- GFP+, ++ M 1 2 3 4 5 6 1 2 3 4 5 6 CAG/CD63-GFP Sry GFP- GFP+, ++ 1 2 3 4 5 6 1 2 3 4 5 6 rat CD63 human CD63 copGFP β-actin

Cultured fibroblast cells from Tg rat Supplementary Figure 2 Cultured fibroblast cells from Tg rat GFP 0 s 2.8 s 5.6 s 8.5 s 11.3 s 14.1 s GFP/DAPI BF

Supplementary Figure 3 Serum EVs (Tg) a Serum EVs (Wt) b c e d anti-human CD63 (1:200) anti-human CD63 (1:50) no first antibody e d

(anti-copGFP/human CD63) Supplementary Figure 4 DIC copGFP copGFP/Hoechst Merged Tg EVs (anti-copGFP) DIC human CD63 human CD63/Hoechst Merged Tg EVs (anti-human CD63) copGFP/human CD63 /Hoechst DIC copGFP human CD63 Merged (anti-copGFP/human CD63) Tg EVs

Supplementary Figure 5 a b Hep cell REF cell Hep EVs rat CD63 Alix Wt Tg Wt Tg Hep EVs Wt Tg rat CD63 Alix TSG101 copGFP Flotillin-1 REF EVs Wt Tg Flotillin-1 copGFP rat CD63 Alix TSG101 LAMP1 (N.D.) Lysate (10 g) human CD63 copGFP rat CD63 Flotillin-1 Alix TSG101 LAMP1