Effect of PTH1-34 on expression of markers for osteoblastic bone formation in <i>Bmi-1^-/-</i> mice.

<p>(A–B) Real-time RT–PCR was performed on humerus extracts from 4-week-old vehicle-treated wild-type (WT) and <i>Bmi-1^-/-</i> mice (KO) and PTH1-34 treated <i>Bmi-1^-/-</i> mice (KO+PTH) for determining the expression of (A) alkaline phosphatase (ALP) and (B) osteocalcin. The expression is calculated as a ratio to the GAPDH mRNA level and shown relative to the levels in vehicle-treated WT mice. (C) Western blots of femur extracts from 4-week-old vehicle-treated WT and <i>Bmi-1^-/-</i> mice and PTH1-34-treated <i>Bmi-1^-/-</i> mice for expression of Runx2, PTHR and IGF-1. β-actin was used as loading control for Western blots. (D-F) Runx2, PTHR and IGF-1 protein levels relative to the β-actin level were assessed by densitometric analysis and presented relative to the levels in vehicle-treated WT mice. For each genotype, n = 6; *: <i>p</i><0.05, **: <i>p</i><0.01, ***: <i>p</i><0.001, compared to vehicle-treated WT mice; #: <i>p</i><0.05, ##: <i>p</i><0.01, ###: <i>p</i><0.001 compared to vehicle-treated <i>Bmi-1^-/-</i> mice.