HG5LN were generated by transfection of cervical cancer HeLa cells with the (GAL4RE)5-betaGlobin-Luciferase-SV40-Neomycin plasmid. rnHG5LN GAL4-rTRalpha cells were generated by transfection of HG5LN cells with the plasmid pSG5-GAL4-rat TR alpha-puromycin.
The 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. rnThe pSG5-puro plasmid pSG5-GAL4-rTR alpha-puromycin enables to express the DNA binding domain of the yeast activator GAL4 (GAL4 DBD) followed by the ligand binding domain of the rat thyroid receptor alpha (rTR alpha LBD). Puromycin N-acetyl transferase selection marker expression confers resistance to puromycin.