Figure 5.

Cellular uptake by HeLa cells after transfection with the TC-1-12-based Cy5-labeled mRNA lipoplexes. Cy5-labeled mRNA lipoplexes were prepared at charge ratios of 3:1 and 4:1 using the MEI and TFH methods, respectively, and added to HeLa cells at 0.5 μg/mL mRNA. As controls, Cy5-labeled mRNA solution (free mRNA) and Lipofectamine MessengerMAX mRNA lipoplexes were added to the HeLa cells. In (A), localization of Cy5-labeled mRNA (green) was observed 3 h after incubation. Scale bar, 100 μm. In (B), geo-mean fluorescence intensity of Cy5-labeled mRNA was measured using flow cytometry 3 h after incubation. Each column represents the mean ± standard deviation (n=3). ^***P<0.001 vs. MEI 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; Cy5, cyanine 5; MEI, modified ethanol injection; TFH, thin-film hydration.