Journal: Pathogens

Article Title: Velocity Gradient Separation Reveals a New Extracellular Vesicle Population Enriched in miR-155 and Mitochondrial DNA

doi: 10.3390/pathogens10050526

Figure Lengend Snippet: Comparison of HIV-negative subjects and HIV-1 patients in terms of plasma EVs miR-92, miR-155, and miR-223 distributions in the iodixanol velocity gradient. Plasmas were pooled ( n = 8 for each group) and pre-treated with proteinase K before EVs precipitation with ExoQuick™. MicroRNA in each gradient fraction was amplified by RT-PCR. Values are normalized as 38—Ct. HIV-1 negative versus ART-naïve ( A , C , E ) and HIV-1 negative versus ART-suppressed and elite controllers ( B , D , F ) were compared using two-way ANOVA and the Bonferroni post-test. Analyses were repeated at least two times (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Purification using the Total Exosomes Isolation kit (Thermo Scientific, Rockford, IL, USA) comprised treating 250 µL of plasma with 50 μL of proteinase K solution (optional, indicated in the Legends), adding 150 μL of precipitation reagent, holding at room temperature for 10 min, centrifuged for 5 min at 10,000× g , resuspending the precipitate in 250 μL of microfiltered PBS.

Techniques: Amplification, Reverse Transcription Polymerase Chain Reaction