Figure 5.
Figure 5. The phosphatidylinositol 3-kinase inhibitor LY294 002 inhibited NF-κB promoter activity and the production of VEGF in Raji cells transiently expressing KSHV G protein–coupled receptor (vGPCR), but only inhibition of VEGF production in K1-transfected cells. Raji cells were transfected with 10 μg pSG5-K1 or vGPCR-pSG5 and cotransfected with 5 μg of the NF-κB–luciferase reporter construct. After 20 hours, the cells were treated with LY294002 (50 μM) for 4 hours and harvested for luciferase activity in cell extracts (□) and VEGF production in culture supernatants (▪) as described in “Materials and methods.” Luciferase values are expressed as a percentage of the value observed in the absence of LY294002 in K1-transfected cells (bar 1). Results represent the means (± SD) of 3 experiments. There was a statistically significant decrease in NF-κB promoter activity in vGPCR-transfected cells (*P < .001, compare bars 5 and 7), but not in K1-transfected cells (P = .3, compare bars 1 and 3). On the other hand, statistically significant decreases were observed in the production of VEGF in cells transfected with K1 (†P < .001, compare bars 2 and 4) and with vGPCR (†P < .001, compare bars 6 and 8).

The phosphatidylinositol 3-kinase inhibitor LY294 002 inhibited NF-κB promoter activity and the production of VEGF in Raji cells transiently expressing KSHV G protein–coupled receptor (vGPCR), but only inhibition of VEGF production in K1-transfected cells. Raji cells were transfected with 10 μg pSG5-K1 or vGPCR-pSG5 and cotransfected with 5 μg of the NF-κB–luciferase reporter construct. After 20 hours, the cells were treated with LY294002 (50 μM) for 4 hours and harvested for luciferase activity in cell extracts (□) and VEGF production in culture supernatants (▪) as described in “Materials and methods.” Luciferase values are expressed as a percentage of the value observed in the absence of LY294002 in K1-transfected cells (bar 1). Results represent the means (± SD) of 3 experiments. There was a statistically significant decrease in NF-κB promoter activity in vGPCR-transfected cells (*P < .001, compare bars 5 and 7), but not in K1-transfected cells (P = .3, compare bars 1 and 3). On the other hand, statistically significant decreases were observed in the production of VEGF in cells transfected with K1 (†P < .001, compare bars 2 and 4) and with vGPCR (†P < .001, compare bars 6 and 8).

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