Effect of Akt knockdown on Stx1-induced production of soluble cytokines. Differentiated THP-1 cells were transfected with Akt-specific siRNA or a nonspecific siRNA control for 48 h. Cells were stimulated with Stx1 for 24 h. (A) Cell lysates were prepared and Western blotting (WB) performed using total Akt antibody. Blots were stripped and reprobed with antibody recognizing actin for equal protein loading. Control (UT), untransfected cells; Control siRNA, nonspecific siRNA. (B and C) Cell supernatants were collected, and soluble TNF-α and IL-1β were quantified by ELISA. All values are expressed in pg/ml ± SEM. Asterisks (*) represent significant differences (P < 0.05) between results with Stx1 alone versus those with Stx1 plus Akt siRNA transfection. RC, reagent control; Con. Si RNA, nonspecific siRNA.