Fig. 2: Superior potency of LNP X is not explained by differences in endosomal escape.
From: Efficient mRNA delivery to resting T cells to reverse HIV latency
a The SNAPswitch assay simultaneously tracks the degree of nanoparticle association, endosomal escape and mRNA expression of LNPs co-encapsulating an AF488 oligo, Cy5-SNAPswitch oligo and mRNA encoding mScarlet. Figure adapted from Liu et al.33 and created in BioRender: Cevaal, P. (2025) https://BioRender.com/3ajxnam. b–g SNAPswitch-reporter Jurkat T cells were incubated with LNPs containing SM-102, DSPC, DMG-PEG2000 and either Cholesterol (Chol) or ß-sitosterol (Sito; LNP X) for 4 h. LNPs encapsulated a reporter mScarlet mRNA, AF488-tagged oligo and a Cy5-SNAPswitch oligo or constitutively active Cy5-oligo. Fluorescence was determined using flow cytometry. b Protein expression of the mScarlet reporter mRNA. c LNP association as determined by the fluorescence of the AF488-tagged oligo. d mScarlet protein expression (as in b) relative to LNP association (as in c). e Amount of cytosolic delivery of nucleic acid cargo as quantified by the Cy5-SNAPswitch oligo fluorescence. f Efficiency of endosomal escape based on the Cy5-SNAPswitch oligo fluorescence (as in e) normalised to a constitutively fluorescent Cy5-tagged oligo. g mScarlet protein expression (as in b) relative to the amount of cytosolic cargo delivery (as in e). b–g Bars represent aggregate mean of n = 3 independent experiments, each symbol representing the average of triplicate technical replicates. Significance was determined using a one-tailed paired ratio t test in (b–e,g) to allow comparisons of MFI values between experiments, or one-tailed unpaired t test in (f), ns non-significant.
