Vasa protein dynamics calculation

Ana Fernandez-Nicolas; Alicia Uchida; Jessica Poon; Mamiko Yajima

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Vasa protein dynamics calculation

AF Ana Fernandez-Nicolas

AU Alicia Uchida

JP Jessica Poon

MY Mamiko Yajima

This method is extracted from research article: Nat Commun, Apr 2022

Vasa nucleates asymmetric translation along the mitotic spindle during unequal cell divisions

DOI: 10.1038/s41467-022-29855-8

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Levels of Kaede-Vasa or Vasa-mCherry-LOV and FIAsH signals were measured on both sides of the mitotic spindle in animal and vegetal blastomeres during the 8~16-cell stage using Fluoview or NIS elements attached to the confocal laser microscopes or by Image J. Regions of interest of the same area on both sides of the spindle and used to measure average fluorescence intensity. This calculation was carried out for each time point and the resulting value divided by average fluorescence intensity immediately after photoconversion or optogenetic activation to determine the change in photoconverted Kaede-Vasa levels or in FIAsH signal over time. Excel was used to create a trendline for photoconverted Kaede-Vasa degradation during anaphase for both sides of the spindle. The slope for each trendline was determined through Excel, and the slope of opposite spindle sides was compared to each other. Each experiment was performed at least three independent times unless individually indicated.

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