Skip to main content
. 2021 Mar 16;22(5):e51475. doi: 10.15252/embr.202051475

Figure EV3. The effect of ALIX‐KD and GW4869 treatment on release of CD63‐positive sEVs and all sEVs.

Figure EV3

  1. MDCK cells stably expressing human CD63 were transfected with siControl or siALIX, and the cells were transferred to cell culture inserts and cultured for 4 days. On the last day, the culture medium was replaced with EV‐depleted medium with or without 10 nM GW4869. sEVs were isolated from the pre‐cleared medium by direct immunoaffinity capture using anti‐CD63 antibody or ultracentrifugation. Cell lysates and sEV samples were analyzed by immunoblotting with the antibodies indicated.
  2. The intensity of the bands shown in (A) was measured in three independent experiments.
  3. sEVs prepared as in (A) were eluted from the beads with a glycine buffer and analyzed by NTA. Representative NTA traces were shown.
  4. Quantification of the NTA data obtained in five independent experiments of (C).

Data information: (B and D) *P < 0.05, **P < 0.01 (one‐way ANOVA and Tukey's test). Mean ± s.e.m. was shown.