Fig. 2

Attenuation of osteogenic transition and osteogenesis-related gene expression in human VICs cultured from FCAVD patients by treatment with ATX inhibitor. A–F Human VICs were cultured in osteogenic differentiation conditions in the presence of BBT-877 or vehicle control. Effects of BBT-877 treatment (1 μM) on ATX activity (A) and LPA species production (B) from VICs during osteogenic differentiation were measured as described in the “Methods” section. C Alkaline phosphatase (ALP) staining and ALP activity of these cells upon exposure to the indicated concentrations of BBT-877 (0, 0.1, 1, 10 μM) after 2 weeks of osteogenic stimulation. D Alizarin red (AR) staining and calcium deposition of the VICs in the presence (0.1, 1, 10 μM) or absence of BBT-877 after 3 weeks of osteogenic stimulation. E The mRNA expression levels of ALPL, RUNX2, SP7, and BGLAP in the VICs in the presence (0.1, 1, 10 μM) or absence of BBT-877. F The mRNA expression levels of FN1, ITGB1, ACTA2, and COL1A1 in VICs in the presence (0.1, 1, 10 μM) or absence of BBT-877. Data are presented as the mean ± SD of triplicates, and a representative set of findings from more than three independent experiments is presented. *P < 0.05, **P < 0.01, ***P < 0.001 versus vehicle control. P values were obtained using a two-tailed t-test