Figure 5.

Impaired osteochondrogenic differentiation in Wnt1^cre;Tfrc^f/f mutants. (a) Expression levels of chondrocyte and osteoblast genes in mandible tissue isolated from E13.5 Wnt1^cre;Tfrc^f/f mutants and controls. Data shown are normalized ratio of Wnt1^cre;Tfrc^f/f /Control (mean±S.E.M); student's t-test; *P<0.05; **P<0.01; ***P<0.001; n≥3. (b) Expression levels of chondrocyte and osteoblast genes in cultured primary mandible mesenchymal cells. Mandible mesenchymal cells were cultured and induced to undergo osteochondrogenic differentiation then collected samples at day 7. Data shown are normalized ratio of Wnt1^cre;Tfrc^f/f/Control (mean±S.E.M); student's t-test; **P<0.01; ***P<0.001; n=5. (c–f) Alcian blue (c) and Alizarin red (e) staining show reduced in vitro osteochondrogenic differentiation in Tfrc KO cells. Mandible mesenchymal cells were cultured and induced to undergo osteochondrogenic differentiation, then collected and stained with Alcian blue at day 0, 4, 7 and 14. (d) and (f) quantification of OD 620 nm and OD 450 nm absorbance data from (c and e), respectively. Data shown are mean±S.E.M; student's t-test; ***P<0.001; control, n=23; Wnt1^cre;Tfrc^f/f, n=14