Figure 1

RNA quantity and transfer method influence biocompatibility and immune response. eGFP-mRNA was transferred into PC-12 cells by lipofection (L) (a) and fusion (F) (b) based standard protocols (SP) and analyzed 24 h after transfer by phase contrast and fluorescence microscopy. Transfection efficiency was determined by flow cytometry. Identical analyses with reduced uptake (RU) conditions were performed for lipofection (c) and fusion (d). Analyses of cells as indicated in a to d for cell death and fluorescence intensity are shown in (e). Cells were additionally harvested for mRNA isolation and characterized for IL-6 and eGFP by qRT-PCR (f). eGFP-mRNA was used in a concentration of 4 µg/mL for fusion and 2 µg/mL for lipofection. In all cases, a total of 1 µg was transferred per substrate. n = At least three independent experiments were used for each analysis. p-values: not significant (n.s.): p > 0.05, *: p ≤ 0.05, **: p ≤ 0.01, ***: p ≤ 0.001.