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Fig. 1. Tamoxifen (TM)-dependent and osteoblast-specific Cre activity using
Cre-ERTM mice. (A) Cre-ERTM transgenic
mice mated with ROSA26 Cre reporter mice (R26R). TM was
injected daily into pregnant females for 3 days from E13.5 to E15.5.
Cre-negative and -positive littermates were stained for β-gal at E18.5.
(B) Cre activity in long bones. Forelimbs were stained for β-gal
at E18.5. C, chondrocytes; black arrow, periosteum; red arrow, osteogenic
center. Scale bars: 50 µm. (C) Cre recombination in calvariae at
E15.5, E16.5 and E18.5 in whole head (lateral and overhead views) and
histology of E18.5 calvariae. Broken line, areas of parietal bones (P). Scale
bar: 20 µm. (D) BMPR1A as evaluated by immunohistochemistry using
cKO calvariae at E18.5. BMPR1A, green; DAPI (nuclei), blue. Scale bars: 10
µm. (E) Immunoblotting for phosphorylated Smad 1/5/8 using E18.5
calvariae. The membrane was treated with polyclonal rabbit
anti-phospho-Smad1/5/8 (1:1000) and monoclonal mouse anti-beta-actin (1:2000),
and visualized by ECL.