Figure 1
From: Confounding factors of ultrafiltration and protein analysis in extracellular vesicle research
Comparative analysis of centrifugal filters for protein and particle recovery of EVs in PBS. (a) 2 mL PBS spiked with GFP-positive EVs was centrifuged at 3,000 g. Concentrate and filtrate were collected. Lysates of filter membranes were used for analysing retained proteins. (b) Particle recovery was analyzed using Nanoparticle tracking analysis (n = 3, *p < 0.0001). (c) Protein recovery was assessed based on protein concentration measurements (n = 3, *p < 0.0001). (d) Retained proteins on the filter membrane were quantified (n = 3, *p < 0.0009). (e) Lysates were used for Western blot immunostaining for GFP. Mean values + SD are indicated. Significant differences were calculated using Student’s t-test with RC 10k as reference. Original immunostaining results are shown in Supplementary Fig. 10. Abbreviations: PBS: phosphate buffered saline. GFP: green fluorescent protein. EV: extracellular vesicle. RC: regenerated cellulose. HY: Hydrosart. PES: polyethersulfone. CTA: cellulose triacetate. WB: Western blot. NTA: Nanoparticle tracking analysis. PCM: protein concentration measurement. RT-qPCR: real time quantitative polymerase chain reaction. EM: electron microscopy.
