A comprehensive Schizosaccharomyces pombe atlas of physical transcription factor interactions with proteins and chromatin. Skribbe et al.
Description
Microscopy images (deconvoluted) related to Skribbe, Soneson, Stadler, et al. Two independent microscopy experiments using strains harboring a lacO array close to the tna1+ locus in different genotypes to monitor its positioning within the nucleus relative to the nuclear envelope (Cut11-mCherry) in both wild-type and Nattou KO Schizosaccharomyces pombe cells by expressing lacI-GFP. For sample explanations, please see README file. For raw images (non-deconvoluted), please contact the lead author of the study.
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Steps to reproduce
Liquid cultures of S. pombe cells were grown over-night in rich medium (YES) at 30°C to the logarithmic phase (OD600 approximately 0.6). Prior to imaging, the cells were attached with lectin (Sigma-Aldrich) to glass bottom dishes with a microwell (MatTek). Cells were imaged on a DeltaVision™ Ultra High-Resolution microscope (Cytiva) with an Olympus 60X/1.42, Plan Apo N, UIS2, 1-U2B933 objective. Z-stacks were obtained at focus intervals of 0.25 μm and images were deconvolved with the inbuilt software softWoRx using default settings. Images were randomized before the analysis to avoid bias. For Z-stack processing, the top and bottom three stacks (out of 22 total stacks) were disregarded, and Maximum Intensity Projection was applied. The FiJi/ImageJ software was used to measure the distances between the foci and the nuclear periphery marked by Cut11-mCherry. For zone-based position quantification the nucleus was divided into three concentric zones with equal surface area (assuming a circular shape of the nucleus). For more details, see https://www.biorxiv.org/content/10.1101/2024.08.20.607873v2.