Microscopy image dataset of Cdk2 and Cdk4 activity reporters, phase contrast and histone H2B paired with nuclear masks of EpH4 mammary epithelial cells across variable cell densities and Cdk inhibition treatments

Fluorescence microscopy dataset of EpH4 mammary epithelial cells constitutively expressing three sensors: DHB-Venus (Cdk2 reporter), mCherry-Cdk4-KTR(Cdk4 reporter), and H2B-mTurquoise (nuclear marker). The dataset contains Cdk2 and Cdk4 activity across different cellular contexts, including varying celldensities and targeted pharmacological inhibition. Cell nuclei were segmented from the H2B-mTurquoise channel using a StarDist '2D_versatile_fluo' model, and the resulting label images are provided alongside the raw images. The dataset also includes the phase contrast channel. Dataset directory structureThe data is organized into two primary directories:● tiff_files/: 288 Raw 16-bit multi-channel TIFF images.● label_files/: 288 Deep learning-derived nuclear segmentation masks. File NamingFiles are named according to the convention: {Condition}_{Replicate}_{field_of_view}.tiff (e.g., A_1_12.tiff). Corresponding label images follow the same convention with a nuclei_prefix (e.g., nuclei_A_1_12.tiff). Code Experimental condition Treatment Cell Density A Low density untreated 5,000 cells/cm2 B High density untreated 50,000 cells/cm2 C Control DMSO vehicle 5,000 cells/cm2 D Cdk2 inhibitor III 60 μM 5,000 cells/cm2 E Palbociclib 1 μM 5,000 cells/cm2 F Cdk2 inhibitor III + Palbociclib 60 μM Cdk2i III, 1 μM Palbociclib 5,000 cells/cm2 Each condition includes three biological replicates. For each replicate, four technical replicates (wells) were obtained, with four independent fields of view captured per well,totaling 16 images (fields of view) per condition per replicate. At total, 288 images were obtained with over 118,000 cells MetadataImages were acquired using a Leica DMI8 wide-field fluorescence microscope equipped with a HC PL FLUOTAR L 20×/0.40 DRY objective and a Leica DFC365 FX monochromatic camera. Four channels were acquired sequentially: Phase Contrast, DHB-Venus (YFP cube, 535 nm), mCherry-Cdk4-KTR (TXR cube, 630 nm), and H2B-mTurquoise (CFP cube, 480 nm). The image resolution is 1392 × 1040 pixels with a pixel size of 0.4604 μm. Associate Code https://github.com/brunicardoso/TranslocNC