Fig 2. In vivo transfection efficiency.
(A) fLuc reporter expression. 10 μg of fLuc or GFP mRNA was prepared in Polyplus in vivo-jet RNA in vivo transfection reagent and administered to Ifnar1-/- via IV and IN combination route. 24 hours following transfection, mice were injected IP with D-luciferin and imaged via IVIS 15 minutes later. (B) hACE2 mRNA stability in vivo. Ifnar1-/- mice were transfected with 10 μg of GFP or hACE2 mRNA. 24 hours post transfection, mice were euthanized and liver and lung tissue homogenized in TriReagent RT for RNA extraction. hACE2 mRNA levels were quantified from extracted RNA via qRT-PCR. Statistical significance was determined by Mann-Whitney test (p = 0.03 and p = 0.03, in the lungs and liver, respectively) (C) hACE2 in vivo transfection efficiency, Ifnar1-/- mice were transfected with 10 μg of hACE2 mRNA or vehicle alone. 24 hours post transfection, mice were euthanized and liver and lung tissue were dissociated into single cell suspensions and stained with a human-ACE-2 specific antibody, followed by an AF488-conjugated anti-human secondary antibody. Live cells were analyzed on an Attune focusing flow cytometer and are represented as frequency of hACE2 expressing cells.