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Intracellular dissociation of mRNA from LNPs. ( a ) Representative confocal images of HEK293 cells after treatment <t>with</t> <t>mRNA-LNP</t> for 6 h: dual-labeled mRNA-LNPs: NBD-cholesterol for LNPs (green) and Cy5 for mRNA (red). Nuclei were stained with Hoechst 33,342 (blue). Scale bars, 25 μm. ( b ) Quantification of intracellular LNP and mRNA colocalization based on 7–10 images for each group, expressed as Pearson’s correlation coefficient and analyzed using ImageJ. Data are presented as mean ± SD. *** p < 0.001
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Intracellular dissociation of mRNA from LNPs. ( a ) Representative confocal images of HEK293 cells after treatment <t>with</t> <t>mRNA-LNP</t> for 6 h: dual-labeled mRNA-LNPs: NBD-cholesterol for LNPs (green) and Cy5 for mRNA (red). Nuclei were stained with Hoechst 33,342 (blue). Scale bars, 25 μm. ( b ) Quantification of intracellular LNP and mRNA colocalization based on 7–10 images for each group, expressed as Pearson’s correlation coefficient and analyzed using ImageJ. Data are presented as mean ± SD. *** p < 0.001
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Intracellular dissociation of mRNA from LNPs. ( a ) Representative confocal images of HEK293 cells after treatment <t>with</t> <t>mRNA-LNP</t> for 6 h: dual-labeled mRNA-LNPs: NBD-cholesterol for LNPs (green) and Cy5 for mRNA (red). Nuclei were stained with Hoechst 33,342 (blue). Scale bars, 25 μm. ( b ) Quantification of intracellular LNP and mRNA colocalization based on 7–10 images for each group, expressed as Pearson’s correlation coefficient and analyzed using ImageJ. Data are presented as mean ± SD. *** p < 0.001
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Intracellular dissociation of mRNA from LNPs. ( a ) Representative confocal images of HEK293 cells after treatment <t>with</t> <t>mRNA-LNP</t> for 6 h: dual-labeled mRNA-LNPs: NBD-cholesterol for LNPs (green) and Cy5 for mRNA (red). Nuclei were stained with Hoechst 33,342 (blue). Scale bars, 25 μm. ( b ) Quantification of intracellular LNP and mRNA colocalization based on 7–10 images for each group, expressed as Pearson’s correlation coefficient and analyzed using ImageJ. Data are presented as mean ± SD. *** p < 0.001
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Intracellular dissociation of mRNA from LNPs. ( a ) Representative confocal images of HEK293 cells after treatment <t>with</t> <t>mRNA-LNP</t> for 6 h: dual-labeled mRNA-LNPs: NBD-cholesterol for LNPs (green) and Cy5 for mRNA (red). Nuclei were stained with Hoechst 33,342 (blue). Scale bars, 25 μm. ( b ) Quantification of intracellular LNP and mRNA colocalization based on 7–10 images for each group, expressed as Pearson’s correlation coefficient and analyzed using ImageJ. Data are presented as mean ± SD. *** p < 0.001
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Intracellular dissociation of mRNA from LNPs. ( a ) Representative confocal images of HEK293 cells after treatment <t>with</t> <t>mRNA-LNP</t> for 6 h: dual-labeled mRNA-LNPs: NBD-cholesterol for LNPs (green) and Cy5 for mRNA (red). Nuclei were stained with Hoechst 33,342 (blue). Scale bars, 25 μm. ( b ) Quantification of intracellular LNP and mRNA colocalization based on 7–10 images for each group, expressed as Pearson’s correlation coefficient and analyzed using ImageJ. Data are presented as mean ± SD. *** p < 0.001
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Intracellular dissociation of mRNA from LNPs. ( a ) Representative confocal images of HEK293 cells after treatment <t>with</t> <t>mRNA-LNP</t> for 6 h: dual-labeled mRNA-LNPs: NBD-cholesterol for LNPs (green) and Cy5 for mRNA (red). Nuclei were stained with Hoechst 33,342 (blue). Scale bars, 25 μm. ( b ) Quantification of intracellular LNP and mRNA colocalization based on 7–10 images for each group, expressed as Pearson’s correlation coefficient and analyzed using ImageJ. Data are presented as mean ± SD. *** p < 0.001
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Intracellular dissociation of mRNA from LNPs. ( a ) Representative confocal images of HEK293 cells after treatment <t>with</t> <t>mRNA-LNP</t> for 6 h: dual-labeled mRNA-LNPs: NBD-cholesterol for LNPs (green) and Cy5 for mRNA (red). Nuclei were stained with Hoechst 33,342 (blue). Scale bars, 25 μm. ( b ) Quantification of intracellular LNP and mRNA colocalization based on 7–10 images for each group, expressed as Pearson’s correlation coefficient and analyzed using ImageJ. Data are presented as mean ± SD. *** p < 0.001
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Intracellular dissociation of mRNA from LNPs. ( a ) Representative confocal images of HEK293 cells after treatment with mRNA-LNP for 6 h: dual-labeled mRNA-LNPs: NBD-cholesterol for LNPs (green) and Cy5 for mRNA (red). Nuclei were stained with Hoechst 33,342 (blue). Scale bars, 25 μm. ( b ) Quantification of intracellular LNP and mRNA colocalization based on 7–10 images for each group, expressed as Pearson’s correlation coefficient and analyzed using ImageJ. Data are presented as mean ± SD. *** p < 0.001

Journal: Journal of Nanobiotechnology

Article Title: Mechanistic insights into mRNA–LNP interactions: role of ionizable lipid content in regulating mRNA intracellular release and translation

doi: 10.1186/s12951-026-04383-5

Figure Lengend Snippet: Intracellular dissociation of mRNA from LNPs. ( a ) Representative confocal images of HEK293 cells after treatment with mRNA-LNP for 6 h: dual-labeled mRNA-LNPs: NBD-cholesterol for LNPs (green) and Cy5 for mRNA (red). Nuclei were stained with Hoechst 33,342 (blue). Scale bars, 25 μm. ( b ) Quantification of intracellular LNP and mRNA colocalization based on 7–10 images for each group, expressed as Pearson’s correlation coefficient and analyzed using ImageJ. Data are presented as mean ± SD. *** p < 0.001

Article Snippet: To investigate how ionizable lipid content influences mRNA-LNP characteristics, we started with benchmark formulations based on the clinically approved Moderna and Pfizer/BioNTech mRNA-LNP formulation.

Techniques: Labeling, Staining