Figure 1

Regulation of PVR/CD155 expression on MM cell lines following treatment with NO donor DETA-NO. A) PVR/CD155 surface expression was analyzed by flow cytometry on SKO-007(J3) cells treated with DETA-NO (200 μM) for 48 h. Data are representative of one out of three independent experiments. The grey colored histogram represents basal expression, while thick black colored histogram represents the expression after treatment with DETA-NO. B) The MFI of PVR/CD155 surface expression was calculated based on at least four independent experiments and evaluated by paired Student t test (*P < 0.05). Histograms represent the MFI with specific mAb subtracted from the MFI value of isotype control. These treatments did not affect the cell viability over the time and DETA-NO concentration [200 μM for SKO-007(J3)] chosen for these experiments (as assessed by PI staining, data not shown). C) Real Time PCR analysis of total mRNA obtained from SKO-007(J3) cells, untreated (−) or treated with 200 μM DETA-NO for 24 h as described above. Data, expressed as fold change units, were normalized with β-actin and referred to the untreated cells considered as calibrator and represent the mean of 3 experiments (*P < 0.05). D-H) The MFI of PVR/CD155 surface expression was calculated for U266, OPM-2, ARK, RPMI-8226 and LP1 MM cells, based on at least three independent experiments and evaluated by paired Student t test (*P < 0.05). Histograms represent the MFI with specific mAb subtracted from the MFI value of isotype control. These treatments did not affect the cell viability over the time and DETA-NO concentration [200 μM for U266, 50 μM for OPM-2, 200 μM for ARK, 100 μM for RPMI-8226 and 125 μM for LP1] chosen for these experiments (as assessed by PI staining, data not shown).