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Fig. 5. ß-catenin responsiveness is restricted to a subset of PR-negative
mouse mammary cells, despite transgene expression in all cells. (A)
Double immunofluorescence analysis of transgene detected by 9E10 (green) and
PR expression detected by anti-PR (red) antibodies in a virgin
MMTV-
N89ß-catenin gland. (B) Expression of
conductin-lacZ detected by X-Gal staining (blue, arrowheads)
indicates that only a subset of ductal cells show a transcriptional response
to uniform transgene expression immunohistochemically detected by 9E10 (brown)
in a section of a
PR+/+;conductin+/lacZ;N89ß-catenin
gland. (C) PR immunohistochemistry on X-Gal-stained sections of
PR+/+;conductin+/lacZ;N89ß-catenin
glands demonstrates distinct populations of ß-catenin-responsive and
PR-expressing cells. (D) Carmine and X-Gal-stained wholemount of a
PR+/+;conductin+/lacZ;N89ß-catenin
gland showing conductin-lacZ expression at sites of alveolar
development. (E) Indirect immunofluorescence for detection of PCNA
(green) and PR (red) reveals that proliferating cells are segregated from
PR-expressing cells. (F) Immunohistochemistry for Ki67 on
PR+/+;conductin+/lacZ;N89ß-catenin
gland showing proliferation of one third of conductin-lacZ-expressing
cells (arrows). Adjacent cells that do not express conductin-lacZ
also proliferate. (G) Graphical representation of the percentage of
Ki67-positive cells within PR-positive, conductin-lacZ-positive and
conductin-lacZ-negative populations. Scale bars: 50 µm in A-C,E,F;
0.3 mm in D.
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