Fig. 3: In vitro characterization of BE-based LNPs containing ionizable lipid A2-B13. | Nature Communications

Fig. 3: In vitro characterization of BE-based LNPs containing ionizable lipid A2-B13.

From: Berberine-inspired ionizable lipid for self-structure stabilization and brain targeting delivery of nucleic acid therapeutics

Fig. 3

a Gel retardation assay of BE@siRNA at LNP/siRNA weight ratios from 0.2 to 20 to evaluate complex stability. b Size distribution analysis by dynamic light scattering. c Transmission electron microscopy (TEM) image showing particle morphology of BE@siRNA. Scale bar, 100 nm. d Cellular uptake comparison between BE@FAM-siRNA and MC3@FAM-siRNA by fluorescence microscopy. Scale bar, 25 µm. e High-magnification imaging of subcellular localization of BE@siRNA and MC3@siRNA. FAM channel shows BE@FAM-siRNAs (green). LysoTracker channel shows lysosome (red). Nuclei were counter stained with Hoechst 33342 (blue). Scale bar, 5 µm. f Analysis of BE-mediated endocytosis mechanisms. g, h LNP@DiD penetration across an in vitro blood-brain barrier (BBB) model using transwell assays. LNPs were labeled DiD (red). Nuclei were counter stained with Hoechst 33342 (blue). Scale bar, 1000 µm. Data are presented as mean ± SD (n = 3 independent experiments). Statistical significance was determined by two-tailed unpaired Student’s t test (two groups) or one-way ANOVA with Dunnett’s multiple comparison tests (multiple groups); ***P < 0.001; NS not significant. Data are representative of three (c, d) and two (e) independent experiments with similar results. Source data are provided as a Source Data file.

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