Fig. 3

Evaluation of Inflammatory Response and Delivery Efficiency of Viral and Non-Viral Delivery Vehicles using RAW264.7-IRCs. A Evaluation of viral and non-viral delivery methods using a dual-reporter assay at 24 and 48 h post-delivery. Efficiency of delivery and inflammatory response were determined by using different viral (AAV 1, 2, 5, 8, 9 and Ad5/35) and non-viral (TransIT-X2 and Lipofectamine 2000) delivery methods carrying a GFP payload to RAW264.7-IRCs at timepoints of 24 h and 48 h post-delivery. %GFP + representing efficient delivery and %mRFP1 + representing inflammatory response. Percentages represent the average of 3 biological replicates. Each bar represents the average percentages from 3 biological replicates. TransIT-X2 and Lipofectamine 2000 were only evaluated at 48 h post-transfection per manufacturer protocol. Raw data points can be found in Table S4. B Evaluation of the geometric mean of GFP to mRFP1 at 24 and 48 h post-delivery. The data represented here is the same samples in 3A. Individual data points were plotted for each biological replicate. Raw data points can be found in Table S4. Statistics done using One-Way Anova and Tukey’s Post-Hoc Analysis can be found in Table S5 for 24 h and S6 for 48 h. C Representative cell populations from dual-fluorescence reporter assay. Populations represented here are RAW264.7 cells with no reporter to set the negative control for GFP/mRFP1, non-treated RAW264.7-IRCS (NT) to determine basal mRFP1 expression among reporter cells, and then three representative populations from RAW264.7-IRCs treated with AAV 1, Ad 5/35, and TransIT-X2, respectively. Gating strategy for dual-fluorescence assay can be found in Figure S2