Immunofluorescent imaging of MCF10A cells treated with OSM, IFNG, or EGF, under siRNA knockdown of 14 genes

This dataset contains immunofluorescent imaging of MCF10A mammary epithelial cells subjected to cytokine treatment and siRNA-mediated gene knockdown. MCF10A cells were cultured in DMEM/F12 growth medium supplemented with horse serum, hydrocortisone, cholera toxin, insulin, and EGF, then transitioned to growth factor-free medium prior to treatment. Cells were treated with either Oncostatin M (10 ng/mL), Interferon-gamma plus EGF (IFNG 20 ng/mL + EGF 10 ng/mL), or EGF alone (10 ng/mL). For gene perturbations, siRNA knockdowns targeting 14 candidate regulators were performed using Lipofectamine RNAiMAX-mediated reverse transfection. Knockdown efficency was assessed functionally through phenotypic readouts. Following siRNA transfection and ligand treatment, cells were imaged using the Incucyte S3 microscope for 48 hours, with images acquired every 30 minutes. Fixed-cell immunofluorescent imaging was performed after 48 hours of ligand treatment. Cells were fixed in 4% formaldehyde, permeabilized, and blocked prior to staining with primary conjugated antibodies for B-Catenin and DAPI counterstaining. Imaging was performed using the InCell 6000 platform. This dataset includes raw imaging data cross all conditions (ligand treatments and siRNA knockdowns), enabling the study of cytokine-incuded collective cell migration and phenotypic transitions in mammary epithelial cells. An excel table is included with Well metadata. Live cell imaging is available at the linked related Zenodo repositories.