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Fig. 6. In mice, TGF-ß1 is unable to inhibit ductal growth of
Wnt5a-/- epithelium in vivo and branching of
Wnt5a-/- organoids in vitro. (A)
Carmine-stained BSA-treated and the contralateral TGF-ß-treated wild-type
mammary gland confirms that the TGF-ß1 pellets (P) are functional and
result in degeneration of the end bud and inhibition of ductal elongation.
Insets represent magnified images of the end buds. (B) Carmine-stained
glands reconstituted with Wnt5a-/- epithelium. The left
gland was treated with BSA and the right contralateral gland was treated with
TGF-ß1. TGF-ß is unable to inhibit the end bud in the absence of
Wnt5a. Insets represent magnified images of the end buds. (C) Ki67
staining in the end buds of wild-type mice either treated with BSA or
TGF-ß. Treatment with TGF-ß resulted in reduced staining relative to
BSA-treated glands. (D) Ki67 staining in the end buds of
Wnt5a-/- reconstituted glands treated with BSA or
TGF-ß. Staining was similar in BSA- and TGF-ß-treated end buds.
(E) Phase contrast images of wild-type mammary organoid preparations
untreated or treated with HGF or HGF and TGF-ß1 shows TGF-ß-mediated
inhibition of HGF-induced branching. Three representative images are shown for
each condition. (F) Wnt5a-/- organoids were
untreated or treated with HGF or TGF-ß and HGF. Untreated
Wnt5a-/- cultures demonstrated increased branching
relative to wild-type cultures. TGF-ß failed to inhibit branching in the
absence of Wnt5a. Three representative images are shown for each condition.
(G) Primary branching was quantified in wild-type and
Wnt5a-/- cultures untreated or treated with HGF or
TGF-ß and HGF. In wild-type cultures treatment with TGF-ß resulted
in a statistically significant decrease in branching. TGF-ß did not
inhibit branching in Wnt5a-/- cultures.
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