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

Fig. 4. Lung and mammary gland epithelial cells undergo squamous metaplasia upon tobacco carcinogen DMBA.

Fig. 4

Human primary lung or mammary gland (Mammary Gl.) epithelial cells were treated with dimethyl sulfoxide (CT) or with 1 μg/ml DMBA for 24 h (A, S-G2/M, and yH2AX) or 72 h (A, HSC and involucrin, B, C). A Flow cytometry quantitations for the percent of, from left to right: cells in the S-G2/M phase of the cell cycle, γH2AX positive cells, cells with high light scatter (HSC) typical of squamous differentiation, involucrin positive cells, all relative to CT. B Representative flow cytometry analyses for the differentiation marker involucrin. +INV, positive cells; T0 = untreated cells. C Representative immunofluorescence for keratin K13 (green) upon the indicated treatments. Blue is nuclear DNA by DAPI. Scale bar, 50 μm. Positive cells by flow cytometry were gated according to negative isotype antibody control. Data are mean ± SEM of two or three replicate samples, representative of 2–3 independent experiment from two different human individuals with similar results. ***p ≤ 0.001, **p ≤ 0.01, *p ≤ 0.05.