Expression of vitamin D3 receptor in KS Y-1, HUVEC, and KS tumor tissue.
VDR mRNA in KS Y-1 (A) and in HUVECs (B). Cells were treated with various concentrations of vitamin D3 for 24 hours, and total RNA was extracted. Total RNA (15 μg) was electrophoresed, blotted, and hybridized to32P-labeled full-length human VDR cDNA (upper panels) and β-actin (lower panels), and exposed to x-ray film until signal was detectable (16 hours for VDR and β-actin in panel A, 3 days for VDR in panel B, and 4 hours for β-actin in panel B). Both cell types express the 4.6 kb mRNA for VDR. Immunocytochemical analysis for VDR expression. KS cell line (KS Y-1) and HUVECs were stained for VDR by using monoclonal antibody. KS cells (D) and HUVECs (F) show positive signal, whereas cells stained with isotype-specific antibodies were negative (C and E, KS and HUVEC, respectively). Antibody was used at 1:50 dilution for the KS Y-1 cells and 1:25 for the HUVECs. KS tumor tissue showed strong VDR-specific signal in spindle cells (H), whereas no staining was observed in KS tumor tissue stained with isotype-specific control (G).
