Choanoflagellate transfection illuminates their cell biology and the ancestry of animal septins

David S. Booth; Heather Szmidt-Middleton; Nicole King

The protocol is summarized in Fig. 2 was set to three standard deviations above the background signal such that any signal above 446 the detection limit has less than a 1% chance of arising from random error. The limit of detection 447 was calculated in two different ways. First, the y-axis intercept and standard deviation were 448 . CC-BY-NC-ND 4.0 International license is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/343111 doi: bioRxiv preprint calculated from a standard curve (Harris, 2007) fit to a serial dilution of nanoluc versus 449 luciferase activity (Fig. S3A) The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/343111 doi: bioRxiv preprint measured using lasers and filters for the fluorophores FITC (Green Fluorescence) and PE (Red 481 Fluorescence); untransfected cells form a population along the y=x line of these plots, and the 482 population of transfected cells are skewed along one axis that corresponds to the fluorophore. 483