Fig. 5

Influence of calphostin C, H88, and H7 on the PKC- or PKA-catalyzed activation of NF-κB in a cell-free system

The results of the cell-free NF-κB activation system with 2 µg of cytoplasmic extract from Jurkat cells in the presence of either 0.2 U of purified PKC or 2 units purified PKA (as in Figs 1–3) are shown. The levels of active NF-κB produced in the cell-free system in the presence of different concentrations of calphostin C (A), H88 (B), or H7 (C) were determined by quantitation of active NF-κB-specific autoradiographic signals. Percent inhibition of the PKC- or PKA-catalyzed release of active NF-κB was calculated from the level of active NF-κB produced in the absence of any drug, which was taken as 0% inhibition. Typical results of three experiments conducted with all three drugs and two enzymes is presented.