Journal: The Journal of Steroid Biochemistry and Molecular Biology
Article Title: The PXR is a drug target for chronic inflammatory liver disease
Figure Lengend Snippet: Effect of PXR activators on NF-κB-dependent gene expression in U397-3XκB-LUC cells. (A) Left panel, RT-PCR for PXR and GAPDH in U937-3XκB-LUC cells, Hep-G2 cells and human skin fibroblasts (Skin FB). Each lane is amplified product from the equivalent 50 ng template total RNA per well, 35 PCR cycles). (B) Luciferase activity after 5 h treatment with the indicated compounds (DMSO, 0.5% (v/v); RIF, 20 μM; HYP, 1 μM; METYR, 250 μM; PCN 20 μM; SULF 1 mM; IKK2-In 10 μM; GT, 3 μM) normalised to LPS + DMSO vehicle activity. Data are the mean and standard deviation of 3 determinations from the same experiment, typical of at least 4 separate experiments. *Significantly different ( P > 95%) reporter gene expression versus LPS-induced DMSO control using Student's t -test (two tailed). (C) Trypan blue exclusion. Upper panels, light micrographs of typical views 5 h after treatment with 300 ng/ml LPS and the indicated compound. Lower panel, percentage trypan blue exclusion in U937-3XκB-LUC cells treated with 300 ng/ml LPS and the indicated concentration of compound for 5 h (note, grey bars are the concentrations employed to examine effects on NF-κB activity in B). Data are the mean of 3 separate incubations from the same experiment, typical of 3 separate experiments. Insol, compound became insoluble when added to culture medium. *Significantly different ( P > 95%) trypan blue exclusion versus LPS-induced vehicle control (DMSO) using Student's t -test (two tailed). Similar results were obtained with compounds without the addition of LPS (data not shown).
Article Snippet: Luciferase expression in U937-3XκB-LUC cells stably transfected with luciferase was determined also using the Dual Luc kit.
Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Amplification, Polymerase Chain Reaction, Luciferase, Activity Assay, Standard Deviation, Two Tailed Test, Concentration Assay