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. Author manuscript; available in PMC: 2015 Nov 24.
Published in final edited form as: Cancer Biol Ther. 2008 Nov 12;7(11):1774–1782. doi: 10.4161/cbt.7.11.6781

Figure 3.

Figure 3

Spontaneous apoptosis of MCF10DCIS.com cells is associated with caspase-9 activation and downregulation of Bcl-2 and p53. (A) Whole cell lysates prepared from monolayers of MCF10A, or MCF10DCIS.com cells continuously passaged for 13 (early), 28 (mid) and 44 (late) passages were analyzed by Western blotting for the indicated markers. Levels of activated 17 kDa caspase-3, p53 and Bax to Bcl-2 ratios relative to β-actin loading control are indicated. (B) Immunohistochemical analysis of Bax (a), Bcl-2 (b) and p53 (c and d) in MCF10DCIS.com xenografts. Thin arrows in (a, b and d), indicate comedo cores and the thick block arrow in (c) indicates p53 reactivity in day 12 MCF10DCIS.com solid DCIS lesions. Original magnification X10 (a and b); X40 (c); X20 (d). (C) A typical labeling pattern of MCF10A (a), or MCF10DCIS.com cells continuously passed for 13 (b) or 46 (c) passages with Mitocapture reagent is shown. Short arrows in (a and b) show cells containing punctuate red fluorescence and long arrows in (c) indicate cells with diffuse green cytoplasm. (d) Graphic representation of percent of cells exhibiting punctuate red fluorescence or diffuse green cytoplasm. Values are mean ± S.E. * and **, indicate significant differences in percent of cells containing punctuate red fluorescence or diffuse green cytoplasm between MCF10DCIS.com cells at passages 13 and 46, respectively (p < 0.001). (e) Agarose gel electrophoresis of genomic DNA isolated from MCF10A or MCF10DCIS.com passage 46 cells.