FIG. 6.

Effect of 1,25(OH)₂D3 on mRNA and protein expressions of ECM-associated proteoglycans in HuLM cells. A, C, and E) Equal amounts of each lysate from HuLM cells treated with increasing concentrations of 1,25(OH)₂D3 (0, 1, 10, 100, and 1000 nM) were examined by Western blot analysis using anti-fibromodulin (A), anti-biglycan (C), and anti-versican (E) antibodies. Western blot analysis with anti-β-actin antibody was used as a loading control. The intensity of each protein band was quantified and normalized to the corresponding β-actin, and then the signal ratio for individual proteins was used to generate the graphs. B, D, and F) Real-time PCR analysis for mRNA levels of fibromodulin, biglycan, and versican genes was performed using gene-specific sense and antisense primers as described in Materials and Methods. The mRNA levels of fibromodulin, biglycan, and versican were normalized to the glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and the normalized values were used to generate the graphs. *P < 0.05 when compared to control, **P < 0.05 when compared between 1,25(OH)₂D3-treated data points. Data are presented as the mean ± SD (n = 3 independent measurements).