H3K27M Nuclei Imaging and Quantification
This data set contains six separate image libraries, each pertaining to an experiment where HEK293 cells were imaged on a confocal microscope to compare the nuclear sizes of either H3K27M and H3-WT, or HDACi treated and untreated cells. Manual size measurement was conducted on thirty randomly selected cells in each experiment group, three repeats for each condition, for a total of ninety selected cells from each library. The attached library includes the size calculations as done in Excel, which shows the meaningful size difference. The images also allow one to appreciate the difference by sight. One of the folders also contains JPEGs of the images selected for the paper in both their original as well as their cropped version, which also includes a scale bar, as well as references to the specific nd2 files from which said JPEGs were created.
Steps to reproduce
HEK293 stably expressing a doxy-inducible promoter were seeded on glass coverslips and induced to express either H3K27M or H3-WT for 4-7 days, or were treated for 72hrs with the HDACi vorinostat at 1uM. The cells were fixed with 4% formaldehyde and stained according to standard protocols. DAPI was used to stain the nuclei. Refer to the methods section in Harpaz, Mittelman, et al., 2022 for more detailed information.