Figure 4

AMPK-α1-knockdown THP-1 cells exhibit a marked reduction in NF-κB-dependent genes in response to LPS stimulation. (A) The wt THP-1 and AMPK-α1^KD THP-1 cells were treated with or without LPS (100 ng/ml) for 3 h. Total RNAs were isolated from each sample and microarray analysis was performed as described in Materials and Methods. The 96 NF-κB-dependent upregulated and downregulated genes were sorted and represented: a, wt THP-1 cells treated with LPS versus wt THP-1 cells; b, AMPK-α1^KD THP-1 cells treated with LPS versus AMPK-α1^KD THP-1 cells; c, AMPK-α1^KD THP-1 cells versus wt THP-1 cells; and d, AMPK-α1^KD THP-1 cells treated with LPS versus wt THP-1 cells treated with LPS. (B) Among 96 NF-κB-dependent genes, 36 genes were further selected and their expression patterns were represented. *, moderately regulated genes; **, highly regulated genes. (C) The WT THP-1 and AMPK-α1^KD THP-1 cells were treated with or without LPS (100 ng/ml) for 3 h. Total RNAs were isolated from each sample and quantitative RT-PCR analysis was performed with specific primers targeted to IL-8, TNF, IL-1β, CD44, and CCL5 genes. Error bars represent mean±S.D. of triplicate samples. (D) The wt THP-1 and AMPK-α1^KD THP-1 cells were treated with or without LPS (100 ng/ml) for 3 h. Total RNAs were isolated from each sample and quantitative RT-PCR analysis was performed with specific primers targeted to NF-KB1, NF-KB1A, NF-KB1E, RELB, IER3, REL, and BCL3 genes. Error bars represent mean±S.D. of triplicate samples