Journal:
Article Title: Functional analysis of peroxisome-proliferator-responsive element motifs in genes of fatty acid-binding proteins
doi: 10.1042/BJ20031340
Figure Lengend Snippet: (A) HepG2 cells were transiently co-transfected with pCAT3 promoter plasmid containing the PPRE, β-Gal, without and with plasmids for PPARγ1 and RXRγ. After 42 h, cells were harvested, and β-Gal and CAT concentrations were determined by ELISAs. For each PPRE, experiments were normalized to those without ectopic PPARγ1/RXRγ, set to unity. Results are means±S.D. for two independent experiments, each performed in triplicate (n=6). (B) HepG2 cells were transiently co-transfected with pCAT3 promoter plasmid containing the PPRE and plasmids for β-Gal, PPARγ1 and RXRγ. DMSO and DMSO plus bezafibrate respectively were added to the medium 4 h after transfection (final concentrations 1% DMSO and 100 μM bezafibrate) and cells were incubated for a further 38 h. After harvest, β-Gal and CAT concentrations were determined by ELISAs. For each PPRE, experiments with ligand were normalized to those with DMSO alone, set to unity. Results are means±S.D. for two independent experiments, each performed in triplicate (n=6).
Article Snippet: Murine promoters of E-FABP gene ( Fabpe [ 22 ]) and H-FABP gene ( Fabph [ 23 ]) respectively served as templates to generate promoter fragments for functional analysis of putative PPREs; Fabpe 2447 (nt −2447 to +35) and Fabph 1514 (nt −1514 to +36) containing the putative PPREs as well as Fabpe 1716 (nt −1716 to +35) and Fabph 817 (nt −817 to +36) without a PPRE were synthesized by PCR and cloned each into the pCAT3 promoter vector (Promega).
Techniques: Transfection, Plasmid Preparation, Incubation
