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. 2021 Apr 13;14(4):356. doi: 10.3390/ph14040356

Figure 2.

Figure 2

Uptake of Cas9 and Cas9-R-phycoerythrin (R-PE)-loaded EVs in Raw 264.7 macrophage cells. MDA-MB-231 human breast cancer cell-derived EVs were loaded with PULSin/Cas9 (Cas9 was labeled with Alexa Fluor 488) or PULSin/Cas9-R-PE. SEC was used to remove free protein, PULSin and PULSin/protein. The Cas9 or Cas9-R-PE amount in each sample loaded through SEC was equivalent and cells were treated with an equal volume of the elution fraction 7–9. Cellular uptake was assessed by flow cytometry after a three-hour incubation period. (a) Uptake of Cas9-loaded EVs and controls. (b) Uptake of R-PE-Cas9-loaded EVs and controls. Data are presented as mean ± SD of three replicates. Statistical analysis was performed by two-way ANOVA with post-hoc pairwise comparisons using Tukey’s test. ****, p < 0.0001 in comparison to PULSin/Cas9 (SEC) or Cas9 (SEC). In figure a, **** pertains to all doses except 0 EVs/cell.