HG5LN were generated by transfection of cervical cancer HeLa cells with the (GAL4RE)5-betaGlobin-Luciferase-SV40-Neomycin plasmid. rnHG5LN GAL4-hPPAR alpha cells were generated by transfection of HG5LN cells with the plasmid pSG5-GAL4-human PPAR alpha-puromycin. rnThe plasmid (GAL4RE)5-betaGlob-Luc-SVNeo contains a luciferase gene driven by a pentamer of yeast activator GAL4 binding sites in front of beta-globin promoter and a neomycin phosphotransferase gene under the control of SV40 promoter.
The pSG5-puro plasmid pSG5-human GAL4-hPPAR alpha-puromycin enables to express the DNA binding domain of the yeast activator GAL4 (GAL4 DBD) followed by the ligand binding domain of the human peroxisome proliferator activated receptor receptor alpha hPPAR alpha LBD. Puromycin N-acetyl transferase selection marker expression confers resistance to puromycin.