Figure 3.

Impact of poly(A) size on eGFP expression in DC2.4 cells

(A) 76A-, 112A-, and 148A-mRNAs were transfected (250 ng mRNA per well, 48-well plate). Fluorescence microscopy analysis was performed 24 h post-transfection. Images were obtained with equal acquisition parameters. Scale bar, 100 μm. (B) Time course analysis of eGFP expression in cells transfected with the different poly(A) mRNAs was performed by flow cytometry. The measurements were acquired at 4, 24, 48, 72, and 144 h post-transfection and MFIs normalized with eGFP expression of 112A-mRNA at 24 h post-transfection. Data were determined as the mean ± SD of 3 independent experiments with four technical replicates. (C) Total eGFP expression of the different mRNAs was represented by calculating the area under the curve (AUC). The relative eGFP expression levels were normalized to that of 112A-mRNA assay. Statistical significance was assessed by nested one-way ANOVA. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗∗p < 0.0001.