Figure 6.
Effect of bafilomycin on EV release measured in the cleared conditioned medium and in the Pellet100K. EVs were collected from DiI-labeled N2a cells treated with Baf (150 nM) or vehicle (UNT) for 4 h and analyzed in the cleared conditioned medium (CCM) or after isolation by UC. The cleared conditioned media were adjusted based on the protein content of donor cells before isolation by UC and analysis by IFC. All samples were counted for the same time (10 min). (A) The total number of DiI-positive EVs in the cleared conditioned medium and in the Pellet100K, respectively, was analyzed. Significance was determined by unpaired t-test and estimation plot analysis performed. The left panel in each plot shows data for individual experiments (circles: untreated; squares: treated with Baf) and their mean. The right panel shows the effect size (difference between means) and its 95% confidence interval. (B) Cleared conditioned medium and Pellet100K preparations were labeled in vitro using anti-CD9 antibodies and Annexin V. Graphs show the relative abundance of differentially labeled EV subpopulations in each fraction. Data are mean percent values ±SD for three independent experiments. Significance determined by unpaired t-test and estimation plot analysis were performed. * p < 0.05.