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( A ) Flow cytometric histogram and MFI analysis of DiI fluorescence from LNPs after coculture with LGG for 6 hours. ( B ) Flow cytometric histogram of FAM fluorescence from miRNA and MFI of FAM fluorescence from miRNA after coculture with LGG for 6 hours. ( C ) Confocal laser scan microscope image of uptake by LGG after coculture for 6 hours. Fluorescence intensity traces (orange line) from the left images were plotted at the right using ImageJ software. ( D ) Flow cytometric histogram and MFI analysis of DiI fluorescence from LNPs after coculture with E. coli (EcN) for 6 hours. ( E ) Flow cytometric histogram of FAM fluorescence from miRNA and MFI analysis of FAM fluorescence from miRNA after coculture with EcN for 6 hours. ( F ) Confocal laser scan microscope image of uptake by EcN after coculture for 6 hours. Fluorescence intensity traces (orange line) from the left images were plotted at the right using ImageJ software. ( G ) The putative binding sites of mdo-miR7267-3p in the monooxygenase ycnE mRNA. ( H ) Quantitative <t>polymerase</t> chain reaction <t>(qPCR)</t> analysis of ycnE expression in LGG treated with mdo-miR7267-3p formulations. ( I ) The pathway of miRNA regulation of indole-3-carboxaldehyde (I3A) production. ( J ) I3A level after cocultured with various mdo-miR7267-3p formulations. Data in (A), (B), (D), (E), (H), and (J) are expressed as means ± SD ( n = 3 biologically independent samples) and representative of two independent experiments. Statistical significance was assessed using one-way ANOVA with Tukey’s multiple comparisons post hoc test, giving P values: * P < 0.05 and *** P < 0.001. PE, phycoerythrin; 5′UTR, 5′ untranslated region; a.u., arbitrary units.
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( A ) Flow cytometric histogram and MFI analysis of DiI fluorescence from LNPs after coculture with LGG for 6 hours. ( B ) Flow cytometric histogram of FAM fluorescence from miRNA and MFI of FAM fluorescence from miRNA after coculture with LGG for 6 hours. ( C ) Confocal laser scan microscope image of uptake by LGG after coculture for 6 hours. Fluorescence intensity traces (orange line) from the left images were plotted at the right using ImageJ software. ( D ) Flow cytometric histogram and MFI analysis of DiI fluorescence from LNPs after coculture with E. coli (EcN) for 6 hours. ( E ) Flow cytometric histogram of FAM fluorescence from miRNA and MFI analysis of FAM fluorescence from miRNA after coculture with EcN for 6 hours. ( F ) Confocal laser scan microscope image of uptake by EcN after coculture for 6 hours. Fluorescence intensity traces (orange line) from the left images were plotted at the right using ImageJ software. ( G ) The putative binding sites of mdo-miR7267-3p in the monooxygenase ycnE mRNA. ( H ) Quantitative <t>polymerase</t> chain reaction <t>(qPCR)</t> analysis of ycnE expression in LGG treated with mdo-miR7267-3p formulations. ( I ) The pathway of miRNA regulation of indole-3-carboxaldehyde (I3A) production. ( J ) I3A level after cocultured with various mdo-miR7267-3p formulations. Data in (A), (B), (D), (E), (H), and (J) are expressed as means ± SD ( n = 3 biologically independent samples) and representative of two independent experiments. Statistical significance was assessed using one-way ANOVA with Tukey’s multiple comparisons post hoc test, giving P values: * P < 0.05 and *** P < 0.001. PE, phycoerythrin; 5′UTR, 5′ untranslated region; a.u., arbitrary units.
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( A ) Flow cytometric histogram and MFI analysis of DiI fluorescence from LNPs after coculture with LGG for 6 hours. ( B ) Flow cytometric histogram of FAM fluorescence from miRNA and MFI of FAM fluorescence from miRNA after coculture with LGG for 6 hours. ( C ) Confocal laser scan microscope image of uptake by LGG after coculture for 6 hours. Fluorescence intensity traces (orange line) from the left images were plotted at the right using ImageJ software. ( D ) Flow cytometric histogram and MFI analysis of DiI fluorescence from LNPs after coculture with E. coli (EcN) for 6 hours. ( E ) Flow cytometric histogram of FAM fluorescence from miRNA and MFI analysis of FAM fluorescence from miRNA after coculture with EcN for 6 hours. ( F ) Confocal laser scan microscope image of uptake by EcN after coculture for 6 hours. Fluorescence intensity traces (orange line) from the left images were plotted at the right using ImageJ software. ( G ) The putative binding sites of mdo-miR7267-3p in the monooxygenase ycnE mRNA. ( H ) Quantitative <t>polymerase</t> chain reaction <t>(qPCR)</t> analysis of ycnE expression in LGG treated with mdo-miR7267-3p formulations. ( I ) The pathway of miRNA regulation of indole-3-carboxaldehyde (I3A) production. ( J ) I3A level after cocultured with various mdo-miR7267-3p formulations. Data in (A), (B), (D), (E), (H), and (J) are expressed as means ± SD ( n = 3 biologically independent samples) and representative of two independent experiments. Statistical significance was assessed using one-way ANOVA with Tukey’s multiple comparisons post hoc test, giving P values: * P < 0.05 and *** P < 0.001. PE, phycoerythrin; 5′UTR, 5′ untranslated region; a.u., arbitrary units.
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( A ) Flow cytometric histogram and MFI analysis of DiI fluorescence from LNPs after coculture with LGG for 6 hours. ( B ) Flow cytometric histogram of FAM fluorescence from miRNA and MFI of FAM fluorescence from miRNA after coculture with LGG for 6 hours. ( C ) Confocal laser scan microscope image of uptake by LGG after coculture for 6 hours. Fluorescence intensity traces (orange line) from the left images were plotted at the right using ImageJ software. ( D ) Flow cytometric histogram and MFI analysis of DiI fluorescence from LNPs after coculture with E. coli (EcN) for 6 hours. ( E ) Flow cytometric histogram of FAM fluorescence from miRNA and MFI analysis of FAM fluorescence from miRNA after coculture with EcN for 6 hours. ( F ) Confocal laser scan microscope image of uptake by EcN after coculture for 6 hours. Fluorescence intensity traces (orange line) from the left images were plotted at the right using ImageJ software. ( G ) The putative binding sites of mdo-miR7267-3p in the monooxygenase ycnE mRNA. ( H ) Quantitative polymerase chain reaction (qPCR) analysis of ycnE expression in LGG treated with mdo-miR7267-3p formulations. ( I ) The pathway of miRNA regulation of indole-3-carboxaldehyde (I3A) production. ( J ) I3A level after cocultured with various mdo-miR7267-3p formulations. Data in (A), (B), (D), (E), (H), and (J) are expressed as means ± SD ( n = 3 biologically independent samples) and representative of two independent experiments. Statistical significance was assessed using one-way ANOVA with Tukey’s multiple comparisons post hoc test, giving P values: * P < 0.05 and *** P < 0.001. PE, phycoerythrin; 5′UTR, 5′ untranslated region; a.u., arbitrary units.

Journal: Science Advances

Article Title: miRNA-loaded biomimetic nanoparticles orchestrate gut microbe to ameliorate inflammatory bowel disease

doi: 10.1126/sciadv.adw5984

Figure Lengend Snippet: ( A ) Flow cytometric histogram and MFI analysis of DiI fluorescence from LNPs after coculture with LGG for 6 hours. ( B ) Flow cytometric histogram of FAM fluorescence from miRNA and MFI of FAM fluorescence from miRNA after coculture with LGG for 6 hours. ( C ) Confocal laser scan microscope image of uptake by LGG after coculture for 6 hours. Fluorescence intensity traces (orange line) from the left images were plotted at the right using ImageJ software. ( D ) Flow cytometric histogram and MFI analysis of DiI fluorescence from LNPs after coculture with E. coli (EcN) for 6 hours. ( E ) Flow cytometric histogram of FAM fluorescence from miRNA and MFI analysis of FAM fluorescence from miRNA after coculture with EcN for 6 hours. ( F ) Confocal laser scan microscope image of uptake by EcN after coculture for 6 hours. Fluorescence intensity traces (orange line) from the left images were plotted at the right using ImageJ software. ( G ) The putative binding sites of mdo-miR7267-3p in the monooxygenase ycnE mRNA. ( H ) Quantitative polymerase chain reaction (qPCR) analysis of ycnE expression in LGG treated with mdo-miR7267-3p formulations. ( I ) The pathway of miRNA regulation of indole-3-carboxaldehyde (I3A) production. ( J ) I3A level after cocultured with various mdo-miR7267-3p formulations. Data in (A), (B), (D), (E), (H), and (J) are expressed as means ± SD ( n = 3 biologically independent samples) and representative of two independent experiments. Statistical significance was assessed using one-way ANOVA with Tukey’s multiple comparisons post hoc test, giving P values: * P < 0.05 and *** P < 0.001. PE, phycoerythrin; 5′UTR, 5′ untranslated region; a.u., arbitrary units.

Article Snippet: RNA extraction kits (catalog no. AG21022), reverse transcription reagent (catalog no. AG11706), and reverse transcription quantitative polymerase chain reaction (qPCR) kits (catalog no. AG11718) were obtained from Accurate Biotechnology (Hunan) Co. Ltd. (Changsha, China).

Techniques: Fluorescence, Microscopy, Software, Binding Assay, Real-time Polymerase Chain Reaction, Expressing