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. 2022 Jun 22;30(9):3034–3051. doi: 10.1016/j.ymthe.2022.06.012

Figure 6.

Figure 6

LNP-siRNA are delivered successfully to mouse MSCs following systemic administration

(A and B) Images depict representative areas of 5-μm bone sections captured 2 days following systemic administration of (A) PBS and (B) DiI-labeled LNP-siRNAs (red), comprising PEG-DSG analyzed by a confocal microscopy. Images demonstrate a degree of overlap between MSCs (green) and LNP-siRNAs (red), indicating the intracellular enrichment of bone MSCs with LNP-siRNAs. Scale bar, 5 μm. The LNP-siRNA delivery to MSCs was also quantified using flow cytometry. (C) The contour plots (top) represent MSCs, defined as PDGFRα+/Sca-1+ cells, at the indicated time points. The contour plots in the bottom panel represent the percentage of DiI-positive MSCs defined as PDGFRα+/Sca-1+ cells of the upper panel, comprising high levels of labeled LNP-siRNAs following their systemic delivery, compared with PBS controls. Plots are representative of two experiments performed in duplicate. (D) Bar graphs demonstrate the percentage of the MSCs (top) containing LNP-siRNAs relative to MSCs of PDGFRα-GFP-expressing mice treated with PBS, and the fold increase of DiI MFI signal in MSCs (bottom) relative to PBS treatments, at the indicated time points. Data from one representative experiment of two independent experiments are shown. Each data point represents arithmetic mean ± SD of three mice per treatment (∗p < 0.001). MFI, mean fluorescence intensity.