Journal: International Journal of Molecular Sciences

Article Title: 3′-UTR Sequence of Exosomal NANOGP8 DNA as an Extracellular Vesicle-Localization Signal

doi: 10.3390/ijms25137294

Figure Lengend Snippet: A graphical depiction of the ratio between the Ct values of EVs and the cytoplasmic 22mer DNA. Ordinary One-way ANOVA was performed for multiple comparisons of the ratio of non-treated sample to 6A, 6B, and the Lipofectamine treated samples. A post hoc Dunnett’s test was used to compare the mean of the non-treated sample ratio to the ratios of the rest of the samples. No treatment vs 6A: Adjusted p value: ****: p < 0.0001, no treatment vs 6B: Adjusted p value: ns: p = 0.1411, and no treatment vs Lipofectamine: Adjusted p value: ns: p = 0.2491. (ns = non-significant).

Article Snippet: Nine days after transfection, i.e., seven days of co-culture and five days of media treatment, the cytoplasmic DNA was extracted using the plasmid DNA extraction kit (Qiagen Catalog number: 27104) following the manufacturer’s protocol.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: 3′-UTR Sequence of Exosomal NANOGP8 DNA as an Extracellular Vesicle-Localization Signal

doi: 10.3390/ijms25137294

Figure Lengend Snippet: A qPCR of HEK293 exosomal, EV, and cytoplasmic DNA. CC: co-culture, MT: media treatment (media from the insert and the well given to naïve cells cultured in a separate plate). Exosome and EV DNA were amplified using an EGFP probe and probe-specific primers. EGFP-specific primers were used for the cytoplasmic DNA amplification. ( A ) CD63 + exosomes, ( B ) no immune separation after PEG-NaCl precipitation from the conditioned media, ( C ) cytoplasmic DNA after co-culture, and ( D ) cytoplasmic DNA after media treatment. Statistical analysis shows no significant difference between the Ct values of EGFP and EGFP-22mer transfected samples. A post hoc Dunnett’s test was used to compare the mean of the EGFP cassette-treated sample with the rest of the samples in the coculture experiments ( A – C ) as well as the media-treatment experiment ( D ). p value explanation is as follows: ( A ) Adjusted p value: *: p = 0.04, ( B ) Adjusted p value: *: p = 0.03, **: p = 0.002, ( C ) Adjusted p value: *: p = 0.02, **: p = 0.007, and ( D ) Adjusted p value: ns: p > 0.1. (ns = non-significant).

Article Snippet: Nine days after transfection, i.e., seven days of co-culture and five days of media treatment, the cytoplasmic DNA was extracted using the plasmid DNA extraction kit (Qiagen Catalog number: 27104) following the manufacturer’s protocol.

Techniques: Co-Culture Assay, Cell Culture, Amplification, Transfection