Fig. 2: Impact of exogenous RNA transfection on global translation in vitro. | npj Vaccines

Fig. 2: Impact of exogenous RNA transfection on global translation in vitro.

From: Synergistic effect of nucleoside modification and ionizable lipid composition on translation and immune responses to mRNA vaccines

Fig. 2

Puromycin incorporation assay to measure global cellular translation by western blot. a Mechanism and workflow for puromycin incorporation assay by western blot. b Western blot image using anti-puromycin (green) and anti-actin (red) antibodies. Cells were transfected with unmodified and modified Sing16 HA mRNA-LNPs over a dose response curve from 1.25–10 µg/million cells. c Quantification % translational inhibition calculated from puromycin signal across the entire lane. Maximum inhibition from Cycloheximide control indicating 100% translational inhibition. Two-way Anova with Šídák’s multiple comparisons test was performed. p > 0.05, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001.

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