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. 2023 Jan 21;9:21. doi: 10.1038/s41420-023-01330-3

Fig. 3. Doxorubicin induces squamous metaplasia in human mammary gland epithelial cells.

Fig. 3

Human primary mammary gland epithelial cells were treated with dimethyl sulfoxide (CT) or with 0.5 μM Doxorubicin (DOXO) for 48 h, as indicated. A Representative flow cytometry analysis for the DNA damage marker γH2AX (+γH2AX, positive cells). The percent of +γH2AX is shown on the right. B Representative flow cytometry analyses of DNA content (2C, 4C, and >4C). The percent of polyploid cells (>4C) is shown on the right. C Representative flow-cytometry analysis for the differentiation marker involucrin (+INV, positive cells). The percent of cells with high light scatter (HSC), or +INV is shown on the right, as indicated. D Top: representative mammary gland colonies after immunofluorescence for keratin K13 (green), treated as indicated. Blue is nuclear DNA by DAPI. Scale bar, 50 μm. Bottom: Orthogonal view profile of a CT mammary gland colony after immunofluorescence for involucrin (INV). E Clonogenic capacity of cells plated after 48 h treatment as indicated and drug-released. Positive cells by flow cytometry were gated according to negative isotype antibody control. Data are mean ± SEM of duplicate or triplicate samples, representative of two independent experiment from two different human individuals with similar results. **p ≤ 0.01, *p ≤ 0.05.