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Fig. 1 | Skeletal Muscle

Fig. 1

From: Isolation of small extracellular vesicles from regenerating muscle tissue using tangential flow filtration and size exclusion chromatography

Fig. 1

Tangential Flow Filtration allows purification of small EVs free of non-EV bound components. A iTEM of anti-HA immunogold labeling after ultracentrifugation protocol from Wnt7a-HA transfected HEK293T cells conditioned media showing isolation of non-EV bound Wnt7a cross-contamination (arrows) with Wnt7a-EVs (arrow heads). B Immunoblot analysis for Wnt7a showing separation of non-EV bound Wnt7a (filtrate) vs. Wnt7a-EVs (retentate) after TFF protocol. Samples were run all in the same gel and have the same time expousure. C iTEM of anti-HA immunogold labeling after TFF protocol from Wnt7a-HA transfected HEK293T cells conditioned media showing the absence of Wnt7a non-EV bound cross-contamination. Membrane bilayer indicated with arrows. D Comparative TEM of EVs isolated with ultracentrifugation and TFF. E Immunoblot analysis comparing EVs isolation efficiency with ultracentrifugation and TFF. F Comparative size distribution analysis of EVs samples after TFF protocol (blue bars) versus classical ultracentrifugation (grey bars). Scale bar 100 nm. Empty plasmid (CTR), Tangential Flow Filtration (TFF), Transmission Electron Microscopy (TEM). (n = 3, mean ± s.e.m., *p < 0.05)

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Skeletal Muscle

ISSN: 2044-5040

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