Figure 7.

LNP delivery of EGFP mRNA transfects human fRPE in vitro. Human fRPE cells were treated with EGFP mRNA encapsulated in 2T LNPs for 2 hours prior to an overnight washout, followed by fixation and immunofluorescence for GFP and ZO-1. Cells were treated with (A) 0.1 mg/mL EGFP mRNA 2T-LNP, (B) 0.5 mg/mL mRNA 2T-LNP, or (C) diluent only (same volume as (A) Images shown are representative of two biological replicates of each treatment (scale bar = 50 µm). (D) Total GFP signal was quantified for fRPE treated with 0.1, 0.25, or 0.5 mg/mL LNP GFP and volume matched diluent or media controls (n = 2 biological replicates per condition). (E) Distribution of segmented GFP signal per cell for each of two biological replicates per treatment (* P < 0.05, ** P < 0.01; nested t-test).