JCS03466 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. (A-D) LacZ staining on mandible bone sections of P1 (A, B) and P10 (C, C) OC-Cre;ROSA26 and control mice. Cre activity was detected in P10 (D) but not in P1 (B) double transgenic mice. (E) The kinetic of Smad4 deletion during the in vitro differentiation of Smad4 mutant primary osteoblasts induced with L-ascorbic acid and β-glycerophosphate. A 234 bp fragment was amplified from the deleted Smad4 alleles with Cre activities. A 481 bp fragment amplified from the Cre transgene was used as the internal control. (F, G) In situ hybridization of distal femurs from 16-day-old control and mutant mice. The expression of PTH/PTHrP receptor (PPR) was decreased in Smad4 mutant (B) compared with control (A).

• Supplemental Figure S2 -

  Fig. S2. The expression of osteoblast markers and molecules in Wnt/β--catenin signaling pathway. (A, B) Real-time PCR of ALP and Runx2 of neonatal calvarial osteoblasts from Smad4 mutant (grey bars) and control mice (white bars). (C-F) Real-time PCR of Lrp5, β--catenin, Lef1 and Dkk2, from calvarial bone extracts of 7-week-old Smad4 mutant (grey bars) and control mice (white bars). *P<0.01.