Fig. 4.

Effects of ABL and TPTD on WNT signaling inhibitors in osteocyte-like cells. a SaOS-2 was cultured under differentiation conditions for 9 or 28 days. Seventy-two hours following the last medium replacement, relative gene expression was measured by qRT-PCR. n = 3–4 replicate wells per group. Data are representative of two independent experiments. b Schema of the experiment regarding (c–f). SaOS-2 was incubated with either ABL or TPTD at a concentration of 100 nmol/L on day 30. Six hours after treatment, the medium was replaced with peptide-free or peptide-containing medium for the rest of the experimental period. c, d Analysis of relative gene expression and protein secretion by qRT-PCR and ELISA under continuous treatment. e, f Analysis of relative gene expression and protein secretion by qRT-PCR and ELISA under transient treatment. *P < 0.05, ***P < 0.001, N.S. not significant. For c to f, n = 3 replicate wells per group. Data are representative of two independent experiments