https://doi.org/10.5061/dryad.sbcc2frfv

Description of the data and file structure

Cell Lines: MCF10A, MCF7 and MDA-MB-231 are commonly cultured cell lines for breast cancer studies. MCF10A is a non-tumorigenic human mammary gland epithelial cell line, which represents normal breast cells. MCF7 is perhaps the most widely studied breast cancer line. MDA-MB-231 is a triple negative breast cancer cell line, often used as a model for breast cancer metastasis. MCF10A was purchased from the American Type Culture Collection (ATCC, Manassas, VA), while MCF7 and MDA-MB-231 cells were gifted to us.

Culture Conditions: MCF10A cells were cultured in MEGM kit (Lonza/Clonetics Corporation) without the GA-1000 (gentamycin-amphotericin B mix) component and with 100 ng/ml cholera toxin. The other two cell lines were cultured in Dulbecco’s Modified Eagle Medium (DMEM) (Sigma) supplemented with 10% EquaFETAL Fetal Bovine Serum (Atlas Biologicals), 100 Units/ml penicillin and 100 μg/ml streptomycin (Fisher Scientific-Hyclone), and were supplemented with 20 mM Hepes (Sigma).

Sample Preparation: Cell lines were maintained under standard culture conditions at 37C and 5% carbon dioxide concentration. Cells were seeded with 7.5 E4 cells/ml density in 6 well plates that had been treated with Fibronectin with density of 2 μg/cm2. Cells were fixed after 22hours.

After fixing, cells were fluorescently labelled with DAPI (Bio legend) for nuclei and with Acti-stain 488 phalloidin (Cytoskeleton, Inc.) for actin, and then imaged using a fluorescent microscope.

Folder contents: This dataset contains 6 folders. The name of the folder denotes the name of the cell line imaged. The appellation A or B refers to a technical replicate. Tiff refers to the format of the image files (tif).

MCF7_A_Tiff

B-labelled technical replicate of the MCF7 cell line.

MCF_7_B_Tiff

B-labelled technical replicate of the MCF7 cell line.

MCF10A_A_Tiff

A-labelled technical replicate of the MCF10A cell line.

MCF10A_B_Tiff

B-labelled technical replicate of the MCF10A cell line.

MDA_MB_231_A_Tiff

A-labelled technical replicate of the MDA-MB-231cell line.

MDA_MB_231_B_Tiff

B-labelled technical replicate of the MDA-MB-231cell line.

Each folder contains a variable number of subfolders. Each subfolder contains three image files in the tif format. The filename with the term DAPI denotes the cell images using the DAPI stain, i.e. the nuclei images. The filename with FITC refers to the cell images with the actin stained as mentioned above. The third file is the composite image.

For example, the subfolder mcf7_a_gda1 can be found in the MCF7_A_Tiff folder. It contains three image files, mcf7_a_gda1_DAPI.tif containing the images of the nuclei, mcf7_a_gda1_FITC.tif containing the image of actin and mcf7_a_gda1_(FITC+DAPI).tif which contains the composite image.