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Figure 2


Fig. 2. PTHrP augments BMP signaling in vivo and in vitro. (A-E) Immunohistochemistry for phospho-SMAD 1, 5, 8 in sections through a PTHrP-/- mammary bud at E15.5 (A), a wild-type bud at E15.5 (B), a PTH1R-/- bud at E15.5 (C), wild-type ventral skin at E18.5 (D) and K14-PTHrP ventral skin at E18.5 (E). Note the nuclear staining for phospho-SMADs in the wild-type mammary mesenchyme (arrows in B) and its absence in the mammary mesenchyme of PTHrP-/- and PTH1R-/- buds (A,C). Also note the induction of nuclear phospho-SMAD staining in the mesenchyme of the ventral skin in K14-PTHrP embryos (arrows in E). (F) Western blot for phospho-SMAD 1, 5, 8 of whole cell lysates from C3H10T1/2 cells treated overnight with graded BMP4 concentrations (0, 10, 50, 100 ng/ml BMP4) with or without 10-7 M PTHrP. The addition of PTHrP augments phospho-SMAD expression (representative blot of five experiments).