Figure 2

Pollutants alone do not affect MCF10A immature features. (a) Bar graph showing cell proliferation of MFC10A cells exposed 60 days to BPA or B(a)P (10⁻¹⁰ M). CT are the control cells (unexposed to pollutants). Data are represented as mean +/− S.E.M., n=21. (b) Column scatter plot showing the number of spheres obtained after 1 week for 1000 seeded cells. One hundred single MFC10A cells exposed 60 days to BPA or B(a)P (10⁻¹⁰ M) were seeded onto 96 ultra-low attachment plates (150 μl medium/well), each value represents the mean of one experiment (mean of 30 wells). CT are the control cells (unexposed to pollutants). Data are represented as mean +/− S.E.M., n=11. **P=0.01 indicates difference between CT and treated cells. (c) After 60 days of exposure to BPA or B(a)P (10⁻¹⁰ M), 200 cells per well were seeded onto 12-well plates in triplicate to perform E-CFC assays for 5–6 days. Bar graph showing the number of different types of E-CFCs obtained per 10 000 seeded cells (mixed colonies, mixed-CFC; myoepithelial colonies, myo-CFC; luminal colonies, lu-CFC). CT are the control cells (unexposed to pollutants). Data are represented as mean +/− S.E.M., n=6. (d,e) Cells were exposed 60 days to BPA (d) or B(a)P (e) (10⁻¹⁰ M), before conducting qPCR analyses to detect the level of expression of differentiation markers. Column scatter plot showing arbitrary units of the ratio to control (CT) cells. Data represent mean +/− S.E.M., n=5. (f) Bright-field images at day 1, 10, 14 or 20 of 3D structures generated in the TDLU assay from control cells (CT), BPA or B(a)P previously exposed (60 days, (10⁻¹⁰ M)) cells. Data are representative of n=15 independent experiments, scale bars=100 μm. (g) Immunohistochemistry of representative TDLU sections, stained for hematoxylin and eosin. Data are representative of n=15 independent experiments, scale bars=100 μm