Fig. 2

1,25(OH)2D3 inhibits cell proliferation by down-regulation of ADAM33 expression. ASM cells were incubated with various doses of 1,25(OH)2D3 for 48 h before treatment or not with 50 ng/ml of VEGF for 30 min, and then cell proliferation was determined by BrdU incorporation (a). ASM cells were incubated at indicated times of 100 nM of 1,25(OH)2D3 before treatment or not with 50 ng/ml of VEGF for 30 min, and then cell proliferation was determined by BrdU incorporation (b). ASM cells were transfected with negative siRNA or ADAM33 siRNA, and then real-time PCR performed. The values are normalized relative to the GAPDH standard (c). ASM cells (d) and ASM cells-ADAM33 (e) were transfected with negative siRNA or ADAM33 siRNA, and then western blotting analysis for ADAM33 was performed. β-actin was used as a loading control. ASM cells-ADAM33 were transfected with negative siRNA or ADAM33 siRNA in the presence of VEGF (50 ng/ml) and 1,25-(OH)2D3 (100 nM) for 48 h, and then cell proliferation was determined by BrdU incorporation (f). All experiments were done at least three times. Values represent the means  ±  SEM. ^* P  <  0.05 vs. control or ASMs-vector; ^# P  <  0.05 vs. VEGF alone or control siRNA or ASMs-control siRNA