Figure 1.

Fabrication and distribution of exosomes and liposomes

(A) Schematic showing protein loading into lung-derived exosomes (RFP-Exos) and liposomes (RFP-Lipos), nebulization administration, ex vivo lung-tissue clearing, and 3D imaging by LSFM. Created with BioRender.com.

(B) TEM images of RFP-Exos and RFP-Lipos; scale bar: 50 nm.

(C) Immunoblot of RFP in exosome and liposome lysate.

(D) Representative immunostaining images of lung parenchymal cells for RFP (red) and DAPI (blue); scale bar: 50 μm.

(E) Quantification of RFP-Exo and RFP-Lipo pixel intensity normalized to nuclei in lung parenchymal cell images; n = 6 per group; data are represented as mean ± standard deviation.

(F) LSFM images of cleared mouse lungs after RFP-Exo and RFP-Lipo nebulization; scale bar: 1,000 μm.

(G) Quantification of the integrated density of RFP normalized to the whole-lung area; n = 74 total slices from two biological replicates per group; data are represented as mean ± standard deviation.

(H) Quantification of the integrated density of RFP normalized to segmented bronchiole and parenchymal regions from whole-lung images; n = 74 total slices from two biological replicates per group; data are represented as mean ± standard deviation.

(I and J) Flow cytometry analysis of lung parenchymal cells co-cultured with RFP-Exos or RFP-Lipos (I) and murine lung cells that received nebulized RFP-Exos or RFP-Lipos (J).