Figure 6.

Impact of different promoters on the regulation of hGLuc expression in various cell types (a) Schematic representation of the Tat-independent, self-inactivating HIV-2 lentiviral vector containing the wild-type or mutant hGLuc gene driven by various internal promoters (Pro): cytomegalovirus (CMV), Rous Sarcoma Virus (RSV), human ubiquitin C (UbC), CMV enhancer SYN hybrid (E/hSYN), human synapsin 1 gene (hSYN) and rat α-tubulin (Tα1). (b–e) Luciferase expression in various cell lines infected with the wild-type or mutant hGLuc containing lentiviral vectors, driven by the various internal promoters. Infected cells were subjected to puromycin to obtain mass populations of transduced cells. Then 2.5 × 10⁵ cells were exposed to 150 µg/ml of G418 or 1 mg/ml of gentamicin for 48 hours before luciferase expression was measured. Wild-type was not exposed to aminoglycoside treatment and luciferase activities are represented in relative light units (RLU). Inductions of mutant hGLuc are represented as fold induction relative to untreated cells. Luciferase readings were normalized to total protein. All experiments were done in triplicate and repeated at least twice. Error bars represent standard deviation.