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. 2022 Feb 16;34(5):1863–1881. doi: 10.1093/plcell/koac043

Figure 1.

Figure 1

Neither trypsin nor RNase A treatment disrupts EVs. A, Protease protection assays of EV cargo proteins PEN1 and TET8. The P40 fractions were subjected to the indicated treatments and then analyzed by immunoblot analysis using anti-PEN1 and anti-TET8 antisera. RNase A and trypsin treatments were performed consecutively in the order indicated for each lane (see “Materials and methods” for details). PEN1 and TET8 were eliminated only when detergent (1% Triton X-100) was included in the mixture, indicating that EVs remained intact when treated with RNase A and trypsin in the absence of detergent. B, Nanoparticle tracking analysis of P40 fractions. Graphs show concentration and size distributions of particles in the three samples. The indicated treatments had no significant effect on these parameters. C, TEM images of negatively stained P40 fractions following the indicated treatments. The bean bag-like particles are EVs. Bars = 0.5 μm.