Figure 4. SMAD4 regulates SHIP1 and IRAK-M during development of endotoxin tolerance.

shSMAD4 and shLUC THP1 cells were treated with 100ng/ml or 10-100ng/ml of LPS for 0, 1, 3, 6 and 24 hours. After the indicated periods, some cells were harvested and lysed for Western blot analysis. A. Culture supernatants assessed for TNF-α concentration. Each treatment was executed in triplicates. Each experiment was repeated at least three times. Results shown are the mean ± SD. B. The cell lysates from the above treatments (A) were subject to Western analysis with indicated antibodies. The blots were reprobed with anti-β-Actin to show equal loading. shSMAD4: shSMAD4 RNA; shLUC: control shRNA.