Figure 5

Examination of osteoblast markers. (a) The Tks4 expression levels were quantified separately in femoral hard bone tissue and BM by RT-PCR. The fold-change differences in the Tks4 mRNA levels were measured in five wild type mice (WT1-WT5), and the fold-changes were calculated using the 2^−ΔΔCt method. (b) BM-MSCs isolated from wild type mice were cultured and differentiated into the osteogenic lineage. Cell lysates were collected during the differentiation period on days 0, 2, 4, 7, 14, and 21, and the Tks4 protein levels were measured using western blotting. Tubulin was used as the loading control. The expression levels of the RunX2 and osteocalcin bone formation markers were measured by RT-PCR separately in BM (c) and hard bone tissue (d). Three 7-month-old mice were used per genotype. The data are presented as the mean ± SEM, and the differences were analyzed by Student’s t-test. ^*p < 0.05.