Supplementary Materials

An RNA biosensor for imaging the first round of translation from single cells to living animals

James M. Halstead, Timothée Lionnet, Johannes H. Wilbertz, Frank Wippich, Anne Ephrussi, Robert H. Singer, Jeffrey A. Chao

Materials/Methods, Supplementary Text, Tables, Figures, and/or References

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  • Materials and Methods
  • Figs. S1 to S12
  • Captions for Movies S1 to S7
  • References

Images, Video, and Other Other Media

Movie S1
Diffusion of mRNA molecules in the cytoplasm. mRNAs were simultaneously imaged in both channels (NLS-PCP-GFP, green and NLS-MCP-TagRFP-T, red). Most mRNAs appear as red particles. A few colocalized particles can be observed. Frame rate: 15ms/frame.
Movie S2
Diffusion of mRNA molecules in the cytoplasm in the presence of CHX (100μg ml-1). mRNAs were simultaneously imaged in both channels (NLS-PCP-GFP, green and NLS-MCP-TagRFP-T, red). Most mRNAs appear as colocalized green/red particles. Frame rate: 15ms/frame.
Movie S3
Diffusion of mRNA molecules in the cytoplasm in the presence of Puromycin (100μg ml-1). mRNAs were simultaneously imaged in both channels (NLS-PCP-GFP, green and NLS-MCP-TagRFP-T, red). Most mRNAs appear as colocalized green/red particles. Frame rate: 15ms/frame.
Movie S4
Diffusion of mRNA molecules in the nucleus. mRNAs were simultaneously imaged in both channels (NLS-PCP-GFP, green and NLS-MCP-TagRFP-T, red). Most mRNAs appear as colocalized green/red particles. Frame rate: 15ms/frame.
Movie S5
Diffusion of mRNA molecules in the nucleus in the presence of low CHX (1μg ml-1). mRNAs were simultaneously imaged in both channels (NLS-PCP-GFP, green and NLS-MCP-TagRFP-T, red). Most mRNAs appear as colocalized green/red particles. Frame rate: 15ms/frame.
Movie S6
Movement of mRNAs molecules in the cytoplasm of arsenite (0.5mM) stressed cells. mRNAs were simultaneously imaged in red (NLS-MCP-Halo, Janelia Fluor 549) and green (NLS-PCP-GFP) channels. P-bodies were imaged in cyan (DDX6-Turquoise2) both before and after mRNA imaging to ensure that they did not move during acquisition of mRNA images. Frame rate: 50ms/frame.
Movie S7
Movement of mRNAs molecules in the cytoplasm of cells relieved from arsenite stress. mRNAs were simultaneously imaged in red (NLS-MCP-Halo, Janelia Fluor 549) and green (NLS-PCP-GFP) channels. P-bodies were imaged in cyan (DDX6-Turquoise2) both before and after mRNA imaging to ensure that they did not move during acquisition of mRNA images. Frame rate: 50ms/frame.