Phosphorylation of CREB upon stimulation of Jurkat cells with EdTx, CT, CyaA, or forskolin.

<p>Cells were stimulated for the indicated times, lysed and processed for western blotting using an antibody against S133-pCREB. On each lane 10 µg of protein were loaded. Upper panel, western blots depicting the time course of EdTx (10 nM EF+40 nM PA), CT (3 nM), CyaA (5 nM), and forskolin (25 µM) dependent CREB phosphorylation. Lower panels, the graphs report the quantitative analysis of the kinetics of CREB phosphorylation induced by EdTx, CT, CyaA, and forskolin (solid line with filled circles). Untreated samples are depicted by the dotted line with black triangles. The intensity of each band was determined using the software Quantity One (Bio-Rad) and was corrected for different loadings using the corresponding value of actin as control. The value at time 0 was set equal to 1. The total amount of cellular CREB did not change during the time course of CREB phosphorylation (not shown). The blots show one representative experiment and the data in the graphs are the mean and standard deviation of three independent experiments.