Fig. 5.

Runx2 regulates the BMP6 autocrine effect on the IP-10/IL-17 co-treatment-initiated OPN/OCN/ALP expression and calcification of HCASMCs. a The HCASMCs were either maintained as the control or were co-treated with IP-10 and IL-17 for 2, 4, 6, and 8 days; b the HCASMCs were either maintained as the control or were pre-treated with IgG or BMP6-blocking antibodies and then co-treated with IP-10 and IL-17 for 8 days; c the HCASMCs were pretreated with smad1- and smad5-specific siRNA and then either maintained as the control or co-treated with IP-10 and IL-17 for 8 days; a–c the expression of runx2 was examined using the western blot test; d–e the HCASMCs were pretreated with control- or runx2-specific siRNA and then maintained as either the control or co-treated with IP-10 and IL-17; d we examined the mRNA expressions of OPN, OCN, and ALP through real-time PCR; e the calcification of the HCASMCs was analyzed using ARS stain. The results in (a–e) are representative of three independent experiments with similar results. The data in (d–e) are mean ± SEM from three independent experiments. * P < 0.05 vs. siCL/control cells; # P < 0.05 vs. siCL/IP-10 and IL-17-co-treated cells