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Relative mRNA abundance measured <t>by</t> <t>RT-qPCR</t> time course of intracellular nuclear mScarlet3 mRNA levels in U-2 OS cells following transfection with constructs 1, 2, 3, 5, 7, and 9 . Data are normalized for each construct. Mean ± SEM from n = 3 independent experiments. B , Live-cell imaging time course of protein production in U-2 OS cells transfected with capped m5C-modified mRNA encoding nuclear-targeted mScarlet3 (orange, top) or mScarlet3-Sec61β (ER marker, Cyan, Bottom) in U-2 OS cells. Representative images shown at indicated timepoints. Scale bars, 100 µm. C , Nuclear mScarlet3 intensity per nucleus following mRNA transfection. The line represents the 95% confidence interval.
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Relative mRNA abundance measured <t>by</t> <t>RT-qPCR</t> time course of intracellular nuclear mScarlet3 mRNA levels in U-2 OS cells following transfection with constructs 1, 2, 3, 5, 7, and 9 . Data are normalized for each construct. Mean ± SEM from n = 3 independent experiments. B , Live-cell imaging time course of protein production in U-2 OS cells transfected with capped m5C-modified mRNA encoding nuclear-targeted mScarlet3 (orange, top) or mScarlet3-Sec61β (ER marker, Cyan, Bottom) in U-2 OS cells. Representative images shown at indicated timepoints. Scale bars, 100 µm. C , Nuclear mScarlet3 intensity per nucleus following mRNA transfection. The line represents the 95% confidence interval.
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Relative mRNA abundance measured <t>by</t> <t>RT-qPCR</t> time course of intracellular nuclear mScarlet3 mRNA levels in U-2 OS cells following transfection with constructs 1, 2, 3, 5, 7, and 9 . Data are normalized for each construct. Mean ± SEM from n = 3 independent experiments. B , Live-cell imaging time course of protein production in U-2 OS cells transfected with capped m5C-modified mRNA encoding nuclear-targeted mScarlet3 (orange, top) or mScarlet3-Sec61β (ER marker, Cyan, Bottom) in U-2 OS cells. Representative images shown at indicated timepoints. Scale bars, 100 µm. C , Nuclear mScarlet3 intensity per nucleus following mRNA transfection. The line represents the 95% confidence interval.
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Relative mRNA abundance measured <t>by</t> <t>RT-qPCR</t> time course of intracellular nuclear mScarlet3 mRNA levels in U-2 OS cells following transfection with constructs 1, 2, 3, 5, 7, and 9 . Data are normalized for each construct. Mean ± SEM from n = 3 independent experiments. B , Live-cell imaging time course of protein production in U-2 OS cells transfected with capped m5C-modified mRNA encoding nuclear-targeted mScarlet3 (orange, top) or mScarlet3-Sec61β (ER marker, Cyan, Bottom) in U-2 OS cells. Representative images shown at indicated timepoints. Scale bars, 100 µm. C , Nuclear mScarlet3 intensity per nucleus following mRNA transfection. The line represents the 95% confidence interval.
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Relative mRNA abundance measured by RT-qPCR time course of intracellular nuclear mScarlet3 mRNA levels in U-2 OS cells following transfection with constructs 1, 2, 3, 5, 7, and 9 . Data are normalized for each construct. Mean ± SEM from n = 3 independent experiments. B , Live-cell imaging time course of protein production in U-2 OS cells transfected with capped m5C-modified mRNA encoding nuclear-targeted mScarlet3 (orange, top) or mScarlet3-Sec61β (ER marker, Cyan, Bottom) in U-2 OS cells. Representative images shown at indicated timepoints. Scale bars, 100 µm. C , Nuclear mScarlet3 intensity per nucleus following mRNA transfection. The line represents the 95% confidence interval.

Journal: bioRxiv

Article Title: mRNAbow : A versatile gene expression system for multiplexed fluorescent imaging using optimized in vitro transcribed mRNA

doi: 10.64898/2026.01.02.697412

Figure Lengend Snippet: Relative mRNA abundance measured by RT-qPCR time course of intracellular nuclear mScarlet3 mRNA levels in U-2 OS cells following transfection with constructs 1, 2, 3, 5, 7, and 9 . Data are normalized for each construct. Mean ± SEM from n = 3 independent experiments. B , Live-cell imaging time course of protein production in U-2 OS cells transfected with capped m5C-modified mRNA encoding nuclear-targeted mScarlet3 (orange, top) or mScarlet3-Sec61β (ER marker, Cyan, Bottom) in U-2 OS cells. Representative images shown at indicated timepoints. Scale bars, 100 µm. C , Nuclear mScarlet3 intensity per nucleus following mRNA transfection. The line represents the 95% confidence interval.

Article Snippet: Quantitative reverse transcription PCR (RT-qPCR) was conducted using the Luna Cell Ready One-Step RT-qPCR Kit (New England Biolabs, E3030) following the manufacturer’s instructions.

Techniques: Quantitative RT-PCR, Transfection, Construct, Live Cell Imaging, Modification, Marker