Luciferase data analysis

Figure 12 In vitro evaluation of the lipidoid library for sIRNA delivery, (a) Percent reduction of firefly luciferase expression in HeLa cells with lipidoid/ siRNA formulations. The data are split into five groupings spanning 0-100% luciferase reduction for ease of analysis, (b) Optimal knockdown levels in HeLa determined by screening at four different lipidoid/siRNA ratios, (c-e) Dose response silencing in HeLa (c), HepG2 (d), and primary macrophage (e) cells. Reprinted from Akinc, A. Zumbuehl, A. Goldberg, M. et at. Nat. Biotechnol. 2008, 26 (5), 561-569, with permission from the Nature Publishing Group.

Figure 5 Quantitative analysis of tripeptide proteasome inhibitor Z-LLL on rAAV-2 transduction in mouse lung. 1x10 viral particles of AV2.RSVluciferase virus was delivered into the mouse lung by nasal inhalation. To evaluate the effects of proteasome inhibitor on gene transfer, 400 (jM Z-LLL (final concentration, representing 1% viral inoculation volume) was coadministrated at the time of viral infection. The entire lung was harvested at 8 months postinfection and luciferase activity was evaluated using a previously described protocol (12). The data represents the mean ( SEM) from four independent mice in each group.

Fig. 2 Increasing content recovery by coupling luciferase-based assays with high-throughput biochemical readouts. The same cell line subjected to the luciferase-based assay protocol (HCT116 cells) is evaluated here for its reliability In reporting a biochemical readout (p53 expression) by dot blot analysis. Cells transfected with indicated expression construct and pathway reporters in a 96-well culture plates were lysed 48 h posttransfection. Following luciferase activity measurements (data not shown), protein from spent lysates was immobilized on nitrocellulose using a liquid handler and filtration manifold, (a) p53 and p-actin protein levels detected using protein-specific antibodies, infrared fluorescent dye-coupled secondary antibodies (with emissions at 680 and 800 nM), and the Li-COR imaging system. Columns of lysate corresponding to cells transfected with p53 DNA are boxed, (b) Quantification of the p53 to p-actin protein ratio...

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