Fluorescent reporters image data: Self-demixing of mRNA copies buffers mRNA:mRNA and mRNA:regulator stoichiometries

Published: 15 November 2023| Version 2 | DOI: 10.17632/5jt3m3twsh.2
Contributors:
Andrés H Cardona, Szilvia Ecsedi, Mokrane Khier, Zhou Yi, Alia Bahri, Amira Ouertani, Florian Valero, Margaux Labrosse, Sami Rouquet, Stéphane Robert, Agnes Loubat, Danielle Adekunle, Arnaud Hubstenberger

Description

Fluorescent reporters raw microscopy data related to Cardona et al., Self-demixing of mRNA copies buffers mRNA:mRNA and mRNA:regulator stoichiometries. This dataset includes fluorescent reporters: - GFP:CAR-1 - SPN-4:GFP Reference Cardona et al., Self-demixing of mRNA copies buffers mRNA:mRNA and mRNA:regulator stoichiometries, Cell (2023), https://doi.org/10.1016/j.cell.2023.08.018

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Steps to reproduce

Live C. elegans nematodes were anesthetized with 50 mM NaN3 in Phosphate Buffer Saline (PBS), and transferred between slides and coverslips to 4% agarose pads, and imaged on a Zeiss LSM 880 microscope with a Plan-Apochromat 40x/1.4 Oil DIC M27 objective and AiryScan SR module. Imaging was conducted in the first 15 min of anesthesia. To avoid signal interference from the intestine, images of GFP-tagged proteins were acquired from the gonad closer to the objective. 3D image data was obtained using Zeiss ZEN software and a 488 nm laser (BP 420-480 + BP 495-550). Z-stacks were acquired with a step-size of 0.5 µm (2.0 optical zoom, 0.66 μs pixel dwell, master gain of 750, digital offset of 1,79 µm pinhole) and processed using the in-built Zeiss Airyscan Processing algorithm.

Institutions

Centre National de la Recherche Scientifique, Universite Cote d'Azur

Categories

Molecular Biology, Cell Biology

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