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Fig. 5. NF-B activation by Tat in ECs. (A) GM7373 were allowed to adhere to plastic coated with the indicated proteins or maintained in suspension (s) for 30 minutes. (B,C) GM7373 adherent to cell culture plates were incubated for 1 hour at 37°C with the indicated concentrations of free GST-Tat (B), or with 100 ng/ml of GST-Tat for the indicated periods of time (C). (D) GM7373 cells adherent to cell culture plates were incubated for 1 hour at 37°C with 100 ng/ml of GST-Tat in the absence (-) or in the presence of SN50, SN50M (both at 25 µg/ml), cRGDfV, cRADfV (both at 3 µM), exoenzyme C3 (5 µg/ml), PP₂ or PP₃ (both at 100 µM). (E) The different clones adherent to cell culture plates were incubated for 1 hour with (black bars) or without (hatched bars) GST-Tat (100 ng/ml). At the end of incubation, cells were lysed and the activation of the indicated NF-B subunit was evaluated. In D, data are expressed as percentage of NF-B activation measured in the absence of any inhibitors. In E, data are expressed as fold increase in respect to basal p50 NF-B activation observed in Tat-untreated cells. In A and D each point is the mean ± s.e.m. of three determinations in duplicate; the data in B, C and E are representative of three independent experiments that gave similar results.

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