FIG. 7.

PI3K/Akt activation by Stx1 results in the activation of the downstream substrate GSK-3α/β. (A) THP-1 cells were stimulated with Stx1, and at each time point, cell lysates were prepared and Western blotting (WB) performed using antibodies specific for phospho-GSK-3α/β. Blots were stripped and reprobed with antibodies recognizing actin for equal protein loading (bottom panel). C, lysates prepared from control cells maintained for 16 h without Stx1. (B) THP-1 cells were treated with the indicated amounts of PI3K (wortmannin) or Akt inhibitors for 1 h prior to stimulation with Stx1 for 1 h (PI3K inhibition) or 4 h (Akt inhibition). Cell lysates were prepared and Western blotting performed using antibodies specific for phospho-GSK-3α/β. Blots were stripped and reprobed with antibodies recognizing actin for equal protein loading (bottom panel). Results shown are representative of at least three independent experiments.