Figure 4.

Effects of the charge ratios of TC-1-12-based mRNA lipoplexes on EGFP levels in HeLa cells transfected with the EGFP mRNA lipoplexes. EGFP mRNA lipoplexes were prepared using the MEI and TFH methods at charge ratios of 2:1 to 5:1, transfected into HeLa cells, and incubated for 24 h. Lipofectamine MessengerMAX was used as a control. (A) EGFP expression in cells was observed using a fluorescence microscope. Scale bar, 100 µm. (B) EGFP transfection efficiency was determined using flow cytometry. Each column represents the mean ± standard deviation (n=3). ^**P<0.01 and ^***P<0.001 vs. TFH method. TC-1-12, 11-((1,3-bis(dodecanoyloxy)-2-((dodecanoyloxy)methyl)propan-2-yl)amino)-N,N,N-trimethyl-11-oxoundecan-1-aminium bromide; EGFP, enhanced green fluorescent protein; MEI, modified ethanol injection; TFH, thin-film hydration.