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. 2020 Dec 14;8:603598. doi: 10.3389/fbioe.2020.603598

Figure 3.

Figure 3

Characterization of EV-depleted FCS (FCSdepl-uc). Prior to ultracentrifugation, normal FCS was diluted in culture medium to different concentrations (20, 50%, and undiluted = 100%); different FCSdepl-uc groups were identified after ultracentrifugation. (A,B) FCSdepl-uc-20% was controlled for EV surface makers (CD9, CD63, and CD81) detected by western blotting (lane 1: hBMSC lysate; lane 2: hBMSC-EVs; lane 3: undepleted FCS; lane 4: FCSdepl-uc-20%). Representative western blot image (A) and Ponceau Red-stained images for each surface marker (B) are shown; n = 3. (C,D) Proliferation and apoptosis of hBMSC were determined by BrdU assay and caspase-3/7 activity assay separately after being incubated in culture medium supplemented with the different FCS groups for 24 h. All values represent mean ± standard deviation. *Significant difference to control: *p < 0.05; **p < 0.01; #Significant difference between groups: #p < 0.05; ###p < 0.001 one-way ANOVA with Newman–Keuls Multiple Comparison Test; n = 4.