DOI: 10.5281/zenodo.13829265
title: Dataset for "The mechanism of nesprin-2 accumulation at the nucleus front during confined cell migration"

description: Data underlying the publication "The mechanism of accumulation of nesprin-2 at the nucleus front during confined cell migration". 

./migrationexperiments/

Folder containing the migration experiment data underlying the main text in the manuscript.It contains the following file and subfolders:



overview.csv: overview file specifying for every cell its identifier (cell number), the cell line, the used transient transfection (if applicable), the types of fluorescence signal, constriction size and acquisition time interval. 

01tiffiles:  folder containing the microscopy tiff files for every cell. The tiff files have been corrected for drift during the acquisition.

02constrictions: folder containing for every cell a file that specifies the position of the constriction center and the tilt angle. The tilt angle is the angle that is needed to tilt the image so that the center of the two pillars forming the constriction are exactly opposite to each other (in all cases, the magnitude of the tilt angle is small (< 0.1 pi)). The coordinate system ranges from (x,y) = (0,0) at the upper left of the tiff image to (x,y) = (xmax,ymax) at the lower right of the tiff image. 

03nucleuscontourcoordinates:  folder containing for every cell a folder that contains for every time step t in the tiff file the coordinates of the nucleus contour. The coordinate system ranges from (x,y) = (0,0) at the upper left of the tiff image to (x,y) = (xmax,ymax) at the lower right of the tiff image. 

04actincontourcoordinates: folder containing the actin contour coordinates for every cell that expressed lifeact-mCherry for every time step at which the full actin contour could be automatically detected. Example reasons why automatic detection did not work are that part of the cell was outside the field of view or the cell was touching another cell. The coordinate system ranges from (x,y) = (0,0) at the upper left of the tiff image to (x,y) = (xmax,ymax) at the lower right of the tiff image. 

05nucleuspositions: folder containing for every cell a file with for every time step in the tiff image, the position of the nucleus center (xn,yn), the position of the nucleus center of mass (xcom,ycom), the position of the nucleus front (xfront, yfront) and the position of the nucleus back (xback, yback). The assignment of what is the front and what is the back of the nucleus is made for the nucleus in the center of the constriction and the same assignment is used throughout the full trajectory. As a result, in the case that a nucleus is rotating during its migration trajectory, the (xback,yback) can be in front of (xfront,yfront) at some time points in the trajectory. The origin (x,y) = (0,0) of the coordinates is at the constriction center. All coordinates have been corrected for tilt angle (as specified in the folder 02constrictions). 

06nucleussizes:  folder containing for every cell a file with for every time step in the tiff image, the x position of the nucleus center (xn), the cross section area of the nucleus as measured from the image and the length of the nucleus contour. The origin (x,y) = (0,0) of the (xn,yn) coordinates is at the constriction center. All (xn,yn) coordinates have been corrected for tilt angle (as specified in the folder 02constrictions). 

07barycenterpositions: folder containing for every cell for which the actin-based cell contour detection worked, the x position of the nucleus center (xn), the x position of the nucleus center of mass (xcom_nucleus), the x position of the cell center of mass (xcom_cell) and the x position of the actin center of mass (xcom_actin). In the case that the actin-based cell contour detection did not work at a certain time step, the xcom_cell and xcom_actin for this time step are indicated with nan (not a number). The origin (x,y) = (0,0) of the coordinates is at the constriction center. All coordinates have been corrected for tilt angle (as specified in the folder 02constrictions). 

08intensityprofiles: folder containing all fluorescence intensity profiles.  The intensity profile files have been sorted in subfolders based on their fluorescence type. The nesprin2 folder contains a separate folder (SUNoverexpression) for the nesprin2 intensity profiles of cells that overexpressed SUN1 or SUN2. The laminAC folder contains a separate folder (mN2Gvariantsoverexpression) for the laminA/C intensity profiles of cells that overexpressed mN2G, mN2G-K274E, mN2G-SR10-13 or mN2G-SR51-54. The origin (x,y) = (0,0) of the (xn,yn) coordinates is at the constriction center. All (xn,yn) coordinates have been corrected for tilt angle (as specified in the folder 02constrictions). 


./sourcedatafigures/

Folder containing the source data for all figure plots in the manuscript (both in the main text and in the supporting information).

 

./plasmidsequences/

Folder containing the DNA sequences of the different mininesprin-2G variants plasmids and the mCherry-SUN1 and mCherry-SUN2 plasmids.