Figure 6.

UTR4 versus UTR2 and 148A-mRNA expression into moDCs and adult zebrafish

UTR2, UTR4, and 148A-mRNAs were transfected into moDCs (125 ng of mRNA per well). (A) Time course analysis of eGFP expression was performed by flow cytometry. The measurements were acquired at 24, 48, and 96 h post-transfection and MFIs normalized with eGFP expression of 148A-mRNA at 24 h post-transfection. Data were determined as the mean ± SD of 3 independent experiments with four technical replicates. (B) 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 148A-mRNA assay. Statistical comparisons were based only on the UTR4 mRNA. Statistical significance was assessed by nested one-way ANOVA. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001. (C) Two or three adult zebrafish were injected intramuscularly with 10 μL of naked mRNA (20 ng/μL) or 10 μL of mRNA-LNP complexes (200 ng of mRNA), respectively. eGFP expression observed with a Leica fluorescent stereoscope on anesthetized animals at 48 h post-injection is shown. The site of injection and the location where the picture was taken (red square) are presented in the drawing. Comparable results were obtained at 24 h post-injection (data not shown). Scale bar, 2 mm.