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. 2016 Jul 12;5(11):1525–1537. doi: 10.5966/sctm.2015-0318

Figure 2.

Figure 2.

Validation of the smRNA system. (A): Time course showing kinetics and stability of MAFA smRNA after single transfection on human pancreatic duct-derived cells (HDDCs) at passages 2–5 (n = 3). (B): Quantitative real-time polymerase chain reaction shows the detection of exogenous PDX1, NGN3, and MAFA smRNAs on HDDCs (passages 2–5) after 7 consecutive daily transfections (n = 5–11). (C): Luciferase assays showed the binding of exogenous MAFA protein to human insulin promoter after transfection on HEK293. MIN6 cells were used as negative control (n = 3–5). (D, E): Fluorescent hybridization probes allowed detection of cytoplasmic MAFA smRNA (red) in 7.6% ± 1.2% of HDDCs after 5 transfections. (F, G): After 5 daily transfections with EGFP smRNA, 47.6% ± 27.4% of HDDCs sustained green fluorescence emission. Scale bars = 400 µm (F), 50 µm (D, G), and 25 µm (E). Abbreviations: eGFP, enhanced green fluorescent protein; MAFA, V-Maf musculoaponeurotic fibrosarcoma oncogene homolog A; smRNA, synthetic modified mRNA.