FIGURE 2.
Assessment of the in vitro transfection efficiency of mRNA mediated by SM102 LNPs. (A) Schematic diagram of SM102 LNPs. (B) Dynamic light scattering distribution plot of SM102 LNPs-Il1rn. (C) Representative transmission electron microscopy image of SM102 LNPs-Il1rn. (D) Denaturing agarose gel electrophoresis of Il1rn mRNA before (lane 1) and after (lane 2) SM102 LNPs encapsulation. (E) Detection of IL-1RA protein in HEK293T cells treated with (lane 2) or without (lane 1) SM102 LNPs- Il1rn mRNA. Lane 3 was the positive control (Commercially available human IL-1RA protein) (F) Representative images showing the SM102 LNPs-mediated transfection of GFP mRNA in HEK293T cells. Scale bar: 50 μm. (G) Flow cytometry results of GFP positive cells for the control group (upper panel) and transfection group (Lower panel) at 48 h post-treatment. (H) Representative images showing the SM102 LNPs-mediated transfection of GFP mRNA in primary tendon stem cells. Scale bar: 50 μm. (I) Quantification of the fluorescence intensity of GFP from 24 h to 72 h post-transfection. (J) ELISA assay for detection of IL-1RA levels in the culture medium of tendon cells treated with 2 ng/μL SM102 LNPs- Il1rn mRNA from Day 1 to Day 7 post-transfection. (K) ELISA assay for detection of IL-1RA levels in the culture medium of tendon cells treated with different concentration of SM102 LNPs- Il1rn mRNA at 24 h post-transfection. *p < 0.05, **p < 0.01, ***p < 0.001; n = 3; All data are shown as the mean ± Standard Error of the Mean (SEM); Statistical significance was determined by one-way ANOVA with Fisher’s LSD test.
