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Figure 4 | BMC Cancer

Figure 4

From: Nitric oxide donors increase PVR/CD155 DNAM-1 ligand expression in multiple myeloma cells: role of DNA damage response activation

Figure 4

NO enhances PVR/CD155 expression: role of DDR. A,B) PVR/CD155 surface expression was analyzed by flow cytometry on SKO-007(J3) cells treated with DETA-NO (200 μM) in the presence or absence of caffeine (CAF 1 mM) or LY294002 (LY 20 μM) for 48 h. Data are representative of one out of four independent experiments. C,D) PVR/CD155 surface expression was analyzed by flow cytometry on SKO-007(J3) cells treated with DETA-NO (200 μM) in the presence or absence of the Chk1/2 inhibitors SB218078 and UCN-01 (0.5 μM and 50 nM respectively) for 48 h. Data are representative of one out of four independent experiments. In these experiments, the concentration used for the different inhibitors, did not significantly affect cell viability as assessed by PI staining (data not shown). E) Real Time PCR analysis of total mRNA obtained from SKO-007(J3) cells, treated for 24 h in the presence or absence of caffeine (1 mM) 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). F) PVR/CD155 surface expression was analyzed by flow cytometry on SKO-007(J3) non-target shRNA (shRNA-control) or pLKO-sh-E2F1 cells, treated with DETA-NO as described above. Data are representative of one out of three independent experiments. G) The MFI of PVR/CD155 surface expression was calculated 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.

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