THP-1 cells stably expressing scrambled, Pellino-1 or Pellino-3 shRNA were differentiated for 72 h with PMA, washed, and pretreated for 20 h with medium or 100 ng/ml LPS. After washing, the cells were treated for 3 h with medium or 100 ng/ml LPS, RNA was prepared, reverse-transcribed, and subjected to RT-qPCR analyses to examine the expression of Pellino-3 (A), TNF-α (B), and HPRT mRNA, using the corresponding gene-specific primers. The data of a representative experiment (n = 3) are depicted. *P < 0.05.