Figure 5. Wnt knockdown impairs osteoblastogenesis.
(A) Alkaline phosphatase expression in RNAs from confluent ST2 cells (Fig. 4A) was analyzed by qPCR and normalized to TBP expression and is presented relative to expression in shControl cells as mean +/− SD of four biological replicates. (B, C) shControl, shWnt6, shWnt10a and shWnt10b ST2 cells were induced to differentiate into osteoblasts. At day 6 post-induction, the extent of osteoblastogenesis was assessed by staining with Alizarin red (B) and by quantification of matrix calcium content (C). Plates and micrographs of Alizarin red-stained cells (B) are representative of three independent experiments. Calcium content (C) was normalized to cellular protein content and is presented as mean +/− SD of four biological replicates per cell type, representative of two experiments. Statistically significant differences in Alp expression or calcium content between shControl and shWnt cells are indicated by *** (= P < 0.001).