Figure 1.

Chloroquine does not substantially affect physical properties of secreted particles. (A) Immunoblot characterization of secreted particles from three cell lines. Protein concentrations of whole cell lysates (WC) and secreted (SE) fractions were quantified using a BCA assay. equal amount of protein was loaded in each lane. Samples from the same experiments were run in parallel blots. Known sEV proteins TSG101, SDCBP/syntenin-1, CD9, and CD63 were analyzed along with known sEV-associated proteins, HSP90AA1 and LAMP2, and the ER marker ERN1/IRE1a. (B) Transmission electron micrograph visualization of sEV contained in the secreted fraction, using uranyl acetate as a counter stain. Scale bar: 200 nm. (C) Total protein content (measured by BCA assay) and particle count (measured by nanoparticle tracking analysis [NTA]) of secreted particles normalized to cell number. significance was determined by paired t-test (**: p < 0.01; n = 3). Horizontal lines denote mean and SD. (D) Size distribution of secreted particles measured by NTA. Solid lines represent mean while dotted lines represent SD (n = 3). (E) Effect of CQ on modal or mean size of secreted particles, or particle/protein ratio. Error bars represent SD. Significance was determined by paired t-test (n = 3).