Multiplexed Atlas of Cytokinesis in Pseudotime

These datasets are the averaged reconstructions of cytokinesis in six steps that were produced by applying Zach Marin's 'Cytokinesis Pseudotime Analysis' package (https://github.com/zacsimile/cytokinesis-pseudotime-analysis) on a collection of 1930 fluorescence spinning disk confocal images. The data are fluorescence images of expanded NRK49F cells, undergoing cytokinesis. The images were acquired as z-stacks with 1 µm spacing and a X/Y-pixel size of 90 nm on a spinning disk confocal microscope. Applying an averaged expansion factor of 4.444, this results in an effective X/Y pixel size of 20.25 nm and 225 nm in Z-direction. These properties can be adjusted for the correct representation of the averaged reconstructions when opening the files in FIJI/ImageJ. The files are composed of 6 time points, representing 6 differet stages of cytokinesis (RC, CS, RS, SM, BA, A). The files are referenced as Supplementary Files 1-4 in the associated preprint. The table below summarizes the file details. z-stack is a volumetric average mean-proj is a 2D mean projected average radial-proj is the axio-radial distribution along the main axis of the cytokinetic bridge File File name Description Supplementary File 1 pseudotime_images_z-stack_reduced_MTs_sen_DAI_Cek_MK1_Ra1_PR1_CiK_ ann_myA_myB_acn_Se7_Se1_Se9_BO4_Tsm_ALm_IS1_CHB_16bit.tif 5D file including all targets that were analyzed and averaged in this study in the 6 cytokinetic stages (order: RC, CS, RS, SM, BA, A) Supplementary File 2 pseudotime_images_mean-proj_MTs_sen_DAI_Cek_MK1_Ra1_PR1_CiK_ann_myA_ myB_acn_Se7_Se1_Se9_BO4_Tsm_ALm_IS1_CHB_16bit.tif 4D file, mean projections of all targets that were analyzed and averaged in this study in the 6 cytokinetic stages (order: RC, CS, RS, SM, BA, A) Supplementary File 3 pseudotime_images_radial-proj_MTs_sen_DAI_Cek_MK1_Ra1_PR1_CiK_ann_myA_ myB_acn_Se7_Se1_Se9_BO4_Tsm_ALm_IS1_CHB_16bit.tif 4D file, axio-radial projections of all targets that were analyzed and averaged in this study in the 6 cytokinetic stages (order: RC, CS, RS, SM, BA, A) Supplementary File 4 pseudotime_images_mean-proj_MTs_sen_DAI_IS1_CHB_16bit.tif 4D file, mean projections of the sub-stages of ESCRT-III cone assembly (order: RC, CS, RS, SM0, SM1, SM2, SM3, BA0, BA1, BA2, BA3, BA4, A) The order of the channels in Suppl. Files 1-3 is as follows MTs tubulin sen Septin2-GFP DAI DAPI Cek CellMask orange MK1 MKLP1 Ra1 MgcRacGAP1 PR1 PRC1 CiK Citron Kinase ann Anillin myA Myosin IIA myB Myosin IIB acn actin Se7 Septin7 Se1 Septin11 Se9 Septin9 BO4 BORG4 Tsm Tsg101 ALm ALIX IS1 IST1 CHB CHMP4B