Figure 3. Centrifugation at 100,000 × g enriches plant EVs much more efficiently than at 40,000 × g.
(A) Schematic of EV isolation by ultracentrifugation of AWF from Arabidopsis. EVs were isolated from clean AWF (isolated by Method 1) via ultracentrifugation at 40,000 × g (P40 fraction) and 100,000 × g (P100 fraction) for 1 hour. For the P100–40 fraction, the supernatant (Sup) of the P40 fraction was centrifuged a second time at 100,000 ×g for 1 hour. (B) Representative TEM images of P100 fraction, P40 fraction and P100–40 fraction isolated from B. cinerea-infected wild-type Arabidopsis. Scale bars, 500 nm. (C) Confocal microscopy of EV fractions (P100, P40 and P100–40) isolated from B. cinerea-infected TET8-GFP/mCherry-PEN1 double-fluorescence transgenic plants. Scale bars, 5 μm.