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. 2022 Dec 16;23(24):16052. doi: 10.3390/ijms232416052

Figure 2.

Figure 2

Characterization of intravesicular RNA and DNA species. (a) Schematic overview of DNase/RNase treatment before sEV lysis and sEV-contained nucleic acid extraction. (b) Nuclease treatment removes non-sEV encapsulated RNA and DNA, but leaves the sEV-membrane intact. Size distribution profiles of sEV samples separated by SEC that were either left untreated or treated with RNase/DNase, obtained by fluorescence NTA measured using a ZetaView TWIN. Samples were stained using the membrane intercalating dye cell mask green. The graph is shown as mean ± SD (n = 12) and is a representative of 3 independent measurements. (c) Concentration of total RNA and DNA in samples without treatment (-) and with RNase/DNase treatment (+) measured using the RNA HS and DNA HS assay, respectively, on a Qubit 2.0 Fluorometer. Data are represented as mean ± SD (N = 3). Statistically significant results after treatment with RNase/DNase are indicated by * for p-value ≤ 0.05 and *** for p-value ≤ 0.001. (d) Ratio of RNA over DNA with RNase/DNase treatment calculated from Figure 1c. (e) Fragment length distribution profiles of samples without treatment and with RNase/DNase treatment obtained using a Bioanalyzer 2100 and RNA 6000 Pico kit (total RNA), small RNA kit (small RNA) or High Sensitivity DNA kit (DNA). Representative profiles of three biological replicate experiments (N = 3) are shown. FU, fluorescence units; nt, nucleotides; bp, base pairs.