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2. Monitoring T cell-dendritic cell interactions in vivo by intercellular enzymatic labelling.
3. Exploiting Secreted Luciferases to Monitor Tumor Progression In Vivo | Springer Nature Experiments
4. Reporter Mouse Models

AB - A straightforward in vivo monitoring technique for biomolecules would be an advantageous approach for understanding their spatiotemporal dynamics in living cells. Abstract A straightforward in vivo monitoring technique for biomolecules would be an advantageous approach for understanding their spatiotemporal dynamics in living cells.

Imaging techniques. Chemical analysis. Chain length. Fatty Acids. Lipid Metabolism. Analytical Chemistry , 86 16 , Cite this: ACS Sens. Article Views Altmetric -. Supporting Information. Cited By. This article has not yet been cited by other publications. Pair your accounts. Your Mendeley pairing has expired. Please reconnect. This website uses cookies to improve your user experience. By continuing to use the site, you are accepting our use of cookies.

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Worms were transfered on cycloheximide-containing plates and incubated for 2 hours, at the growth temperature. After photobleaching, worms were returned in cycloheximide-containing plates during fluorescence recovery. To determine the average and maximum pixel intensity for each image of fluorescent cell or tissue of interest in the collected photomicrographs we processed images acquired in previous steps with the image processing software ImageJ Rasband, W. For each cell, tissue or animal, images were converted to a pixel depth of 8 bit shades of grey. On occasion area continuity, high contrast ratios , selection of the fluorescent area was done automatically.

We adjusted the threshold until the region of interest was marked. Mean values were compared using unpaired t tests. The R2 linear regression tool of the Microsoft Office Excel was used to generate best-fit lines corresponding to radioactive incorporation or fluorescence recovery rate.

Monitoring T cell-dendritic cell interactions in vivo by intercellular enzymatic labelling.

Analysis of variance ANOVA was used for comparisons of multiple groups of values, followed by Bonferroni-corrected multiple-group comparison posthoc t tests. Monitoring of protein synthesis by conventional radioactive metabolic labeling. Incorporation of radioactive amino acids into nascent polypeptides in wild type animals at the indicated time points after growth on radioactive amino acid food source, either in the absence black line or in the presence grey line of the protein synthesis inhibitor cycloheximide.

Fluorescence recovery in wild type animals expressing p pqn PQNGFP, a full-length transcription factor reporter fusion expressed at low levels PQN; zinc-finger family; tight nuclear localization. Regression analysis of fluorescence recovery in wild type animals expressing a full-length p myo- 3 MYOGFP myosin fusion, which localizes in the myofilament lattice, specifically in the body wall muscles. Fluorescence recovery in wild type animals expressing a full-length p mec-4 MECGFP ion channel fusion, which sorts through the Golgi and the endoplasmic reticulum and localizes on the plasma membrane, specifically in the six touch receptor neurons.

Fluorescence recovery in wild type animals expressing either a p myo-3 mitoGFP reporter fusion, localized in mitochondria of body wall muscles grey line or a p asp-4 ASPGFP reporter fusion, localized in lysosomes black line. Regression analysis of fluorescence recovery in wild type animals expressing either a p sod-3 GFP transcriptional reporter fusion amino acids; black line , or a full-length, p ife-2 IFEGFP chimera amino acids; dotted line , or a full-length p clp-1 CLPGFP fusion amino acids; grey line.

We thank Angela Pasparaki for expert technical support and members of our laboratory for comments and suggestions. Some nematode strains were provided by the C. We thank A. Fire for plasmid vectors. Conceived and designed the experiments: NK NT.

Exploiting Secreted Luciferases to Monitor Tumor Progression In Vivo | Springer Nature Experiments

Performed the experiments: NK NT. Analyzed the data: NK NT. Wrote the paper: NK NT. Browse Subject Areas? Click through the PLOS taxonomy to find articles in your field. Abstract Analysis of general and specific protein synthesis provides important information, relevant to cellular physiology and function.

Introduction Proper regulation of protein synthesis is critical for cell growth, cell proliferation and cell death. Results Monitoring de novo protein synthesis by FRAP Conventional FRAP applications usually involve highly localized photobleaching of fluorophores, within defined sub-cellular areas or compartments by means of a laser beam, under a confocal microscope [7] , [8]. Download: PPT. Figure 1.

Comparative analysis of protein synthesis rates between different genetic backgrounds Assessment of protein synthesis rates is an important component of mRNA translation regulation studies. Figure 2. Regression analysis of fluorescence recovery in both wild type and IFE-2 deficient animals expressing p ife-2 GFP throughout somatic tissues. Figure 3. Cell and tissue-specific monitoring of protein synthesis Different cell types and tissues show intrinsically different protein synthesis activities. Figure 4. Regression analysis of fluorescence recovery in cell and tissue-specific level. Figure 5.

Discussion We have developed a non-radioactive and non-invasive approach for monitoring protein synthesis in vivo , based on live imaging of fluorescence recovery after photobleaching.

Reporter Mouse Models

Materials and Methods Nematode strains We followed standard procedures for C. Sample preparation, photobleaching and recovery The FRAP procedure was performed either directly on a plate or on a coverslip. Cycloheximide treatment In order to verify that fluorescence recovery is due to new protein synthesis we used the antibiotic cycloheximide, a potent and specific inhibitor of protein synthesis.

Quantification of GFP emission and protein synthesis rate To determine the average and maximum pixel intensity for each image of fluorescent cell or tissue of interest in the collected photomicrographs we processed images acquired in previous steps with the image processing software ImageJ Rasband, W. Supporting Information. Figure S1. Figure S2.

Figure S3. Figure S4. Figure S5. Figure S6. Materials and Methods S1.

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Acknowledgments We thank Angela Pasparaki for expert technical support and members of our laboratory for comments and suggestions. References 1. Annu Rev Biochem — View Article Google Scholar 2.

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Ibba M, Soll D Quality control mechanisms during translation. Science — View Article Google Scholar 3. Cancer Cell 5: — View Article Google Scholar 4.



Nature — View Article Google Scholar 5. Martin R Protein synthesis : methods and protocols. Totowa, N. Am J Physiol E— View Article Google Scholar 7. Gribbon P, Hardingham TE Macromolecular diffusion of biological polymers measured by confocal fluorescence recovery after photobleaching. Biophys J — View Article Google Scholar 8.