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. Author manuscript; available in PMC: 2021 Nov 1.
Published in final edited form as: J Cell Physiol. 2020 Jan 22;235(11):8210–8223. doi: 10.1002/jcp.29476

Figure 1: Isolation and characterization of EVs isolated from the fetal lung fluids.

Figure 1:

(A) Lung fluid from seven fetuses at E18.5 of gestation were collected and combined. EVs were isolated by ultracentrifugation. EVs concentration was measured using NanoSight. (B) Graphical representation of size distribution of EVs measured by Nanoparticle Tracking Analysis. (C) Detection of EVs was evaluated by Western dot blot using anti-CD63 antibody. Lanes 1 and 2 represent the pellet containing cell debris and apoptotic bodies after 1,000 and 14,000 rpm whereas lane 3 is the pellet after ultracentrifugation containing EVs. (D) Flow cytometry analysis to assess purity of EVs isolation using anti-CD9 and anti-CD63 antibodies. Blue histograms represent tetraspanin-coupled magnetic beads only (described in methods) and in absence of EVs the detection antibody is unable to bind with magnetic beads which indicate the specificity of detection antibodies for purity and integrity of EVs. Pink histogram represents the negative control in absence of detection fluorescence antibody. Yellow histogram represents EVs that bind to tetraspanin-coupled beads and positive for the detection of the fluorescence antibody. Representative figures are shown for at least two independent experiments.