Fig. 2. Optimization of N/P ratio for in vitro transfection by FACS.
GFP mRNA was encapsulated in each LNP and transfected into A549 and BEAS-2B cells to determine the optimal N/P ratio. Encapsulation was performed at room temperature for 30 min at each N/P ratio, followed by transfection into A549 and BEAS-2B cells. After 3 h of transfection, cells were incubated at 37℃ with 5% CO2 for 21 h. Post-trypsinization, cells were collected, and GFP expression efficiency was assessed using a Flow cytometer. (A) Percentage of GFP expression efficiency measured by FACS analysis. (n = 3; mean ± SD, ***P < 0.001 as compared with control; Student's t-test, two-tailed). (B) Confirmation of the shift in GFP expression efficiency measured by FACS analysis, shown in the histogram.
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