Figure 3. Knockdown of SMAD4 by shSMAD4 RNA results in increased p-AKT and p-IκBα levels in THP1 cells.

A. Isolated proteins from shSMAD4 and shLUC THP1 cells were subjected to Western blot analysis with the indicated antibodies. Blots were reprobed with anti-β-Actin to show equal loading. B. shSMAD4 and shLUC THP1 cells were treated with 100ng/ml or 10-100ng/ml of LPS for 0, 10, 30 and 60 minutes. After the indicated periods, cells were harvested for Western blot analysis with indicated antibodies. Blots were reprobed with anti-β-Actin to show equal loading. shSMAD4: shSMAD4 RNA; shLUC: control shRNA. All results are representative of three separate experiments.