Figure 3From: Targeting 3-phosphoinositide-dependent protein kinase 1 by N-acetyl-cysteine through activation of peroxisome proliferators activated receptor alpha in human lung cancer cells, the role of p53 and p65NAC induces PDK1 promoter activity via PPARα. A, The human PDK1 wild type promoter construct schematic is presented. These regions contain several transcription factor binding sites including c-myc, NF-κB, p53, among others. B, A549 and H1792 cells (1 × 105 cells) were cotransfected with a wild type PDK1 promoter construct (shown in A) ligated to a luciferase reporter gene and an internal control phRL-TK Renilla Luciferase Vector for 24 h using the oligofectamine reagent (Invitrogen) according to the manufacturer’s instructions. After 24 h of incubation, cells were treated with NAC (5 mM) and Fenofibrate (10 μM) for an additional 24 h. C, A549 (1 × 105 cells) were cotransfected with a wild type PDK1 promoter construct ligated to a luciferase reporter gene and an internal control phRL-TK Renilla Luciferase Vector for 24 h using the oligofectamine reagent. After 24 h of incubation, cells were treated with GW6470 (20 μM) for 2 h, followed by NAC (5 mM) and Fenofibrate (10 μM) for an additional 24 h. Afterwards, the ratio of firefly luciferase to renilla luciferase activity was quantified.Back to article page